Phytomedicine

Combining cisplatin with Pinellia pedatisecta Schott lipid-soluble extract induces tumor immunogenic cell death in cervical cancer

Pinellia pedatisecta Schott extract (PE) is extracted from Pinellia pedatisecta Schott (PPS), a traditional Chinese medicinal plant with the potential for direct anticancer effects or eliciting an anti-tumor response by activating the immune system. To explore PE's ability and mechanism to reconstruct cisplatin's immunogenicity. Cervical cancer cells were treated with cisplatin (CDDP) and/or PE. The exposure of calreticulin (CRT) on cell membrane was investigated by flow cytometry. The extracellular of ATP and HMGB1 was investigated by Western blot analysis, immunofluorescence and ELISA assay. Changes in immune profiles were using flow cytometry in vaccination and anti-tumor assays in vivo. Lastly, the mechanism of PE influenced the ROS/ERS pathway was examined by ROS assay kit, flow cytometry and Western blotting. PE treatment induced translocation of CRT from the endoplasmic reticulum to the cell membrane of tumor cells, concomitantly triggering immunogenic cell death (ICD). In terms of mechanisms, endoplasmic reticulum (ER) stress relievers could impede the ability of PE to induce immunogenicity. This indicates that PE is activated by ER stress, leading to subsequent induction of ICD. Upon analyzing RNA-seq data, it was observed that PE primarily induces programmed cell death in tumors by impeding upstream antioxidant mechanisms. Additionally, it transforms dying tumor cells into vaccines, activating a series of immune responses. This study observed for the first time that PE-induced CRT exposure on the membrane of cervical cancer cells compensates for the defect of nonimmunogenic cell death inducer CDDP thereby stimulating potent ICD. This ability restores the immunogenicity of CDDP through ER stress induced by the ROS signal. ROS played a role in PE's ability to induce ICD, leading to increased expression of ER stress-related proteins, including ATF3 and IRE-1α. PE exerted anti-cancer effects by increasing the ROS levels, and ROS/ERS signaling may be a potential avenue for cervical cancer treatment. Hence, the synergistic use of PE and CDDP holds potential for enhancing immunochemotherapy in cancer treatment.

Daphnetin induces ferroptosis in ovarian cancer by inhibiting NAD(P)H:Quinone oxidoreductase 1 (NQO1)

Ferroptosis, an emerging nonapoptotic, modulated cell death process characterized by iron accumulation and subsequent lipid peroxidation, has been intimately implicated in the development and progression of ovarian cancer (OC). Daphnetin (Daph), a natural product isolated from Daphne Korean Nakai, exhibits anticancer efficacy against various solid tumors. However, the specific role and potential mechanism underlying Daph-mediated modulation of ferroptosis in OC cells remain elusive. This study aims to analyze the proferroptotic impacts of Daph on OC cells and to further explore the underlying mechanisms involved. We used CCK-8, wound healing and Transwell assays to assess whether Daph can inhibit the proliferation and migration of OC cells. Additionally, transmission electron microscopy (TEM), iron measurement, reactive oxygen species (ROS) analysis, lipid peroxidation assays, qRT-PCR and western blotting were utilized to evaluate the impact of Daph on ferroptosis and elucidate the potential underlying mechanism. Furthermore, RNA sequencing analysis, molecular docking analysis, cellular thermal shift assays (CETSAs) and NQO1 activity assays were used to predict and validate the binding and mechanistic interactions between Daph and NQO1. Subcutaneous tumorigenesis models were utilized to examine the effectiveness of Daph (and/or cisplatin) in vivo. Daph exerted antitumor effects by inducing the death and suppressing the migration of A2780 and SKOV3 cells. Further, Daph induced ferroptosis in OC cells, as evidenced by the accumulation of intracellular ferrous iron (Fe2+), ROS and lipid peroxides, as well as the decreases in the glutathione/oxidized glutathione disulfide (GSH/GSSG) ratio and the expression of ferroptosis indicators (SLC7A11 and GPX4). RNA sequencing and molecular docking analyses revealed that the direct interaction between NQO1 and Daph reduced both the activity and expression of NQO1. Importantly, NQO1 overexpression effectively alleviated the effects of Daph on proliferation, migration, and ferroptosis in vitro and in vivo. Interestingly, we also found that combination treatment with Daph, a negative regulator of NQO1, and cisplatin synergistically induced cytotoxicity in OC cells. Our findings are the firstly demonstrated that Daph acts as a novel ferroptosis inducer in OC cells by specifically targeting NQO1 and is thus a promising candidate agent for OC treatment.

Flavonoid GL-V9 as a novel and potent acetyl-coa carboxylase 1 inhibitor confers cisplatin hypersensitivity in ovarian cancer

Cisplatin has been a cornerstone of Ovarian cancer (OC) treatment since its clinical introduction in the 1970s, with platinum-based regimens forming the core of first-line therapy. However, approximately 70% of patients eventually experience disease recurrence and develop resistance to platinum. To date, few clinically viable alternatives to cisplatin have emerged for the effective management of OC. Therefore, this study aims to identify a novel agent capable of enhancing cisplatin sensitivity and overcoming chemoresistance in ovarian cancer. This study evaluates the anti-tumor effects of GL-V9 in ovarian cancer, elucidates its underlying mechanisms, and evaluates its potential for clinical translation. The mechanisms underlying the effects of GL-V9 were investigated using streptavidin pull-down assays, functional analyses (cell viability, clonogenic survival, and apoptosis), and Western blotting. The antitumor efficacy of GL-V9 alone and in combination with cisplatin was further evaluated in an A2780 ovarian cancer xenograft model. We found that GL-V9, a flavonoid derived from Scutellaria, exerted potent antitumor effects in OC by inducing apoptosis and inhibiting proliferation. Furthermore, GL-V9 enhanced cisplatin sensitivity and reversed cisplatin resistance. Mechanistically, GL-V9 directly bound to and inhibited acetyl-CoA carboxylase 1 (ACC1), leading to elevated intracellular reactive oxygen species (ROS) levels. Both in vitro and in vivo experiments confirmed that GL-V9, alone or in combination with cisplatin, significantly suppressed tumor growth without observable toxicity. GL-V9 overcomes cisplatin resistance in ovarian cancer by targeting ACC1 and elevating ROS levels, highlighting its potential as a clinically translatable therapeutic strategy.

Daphnetin triggers ROS-induced cell death and induces cytoprotective autophagy by modulating the AMPK/Akt/mTOR pathway in ovarian cancer

Ovarian cancer is one of the most common gynecological malignancies in the world. Daphnetin (Daph) was previously reported to possess antitumor potential, but its potential and molecular mechanisms in ovarian cancer remain poorly understood. In the current study, we aimed to explore the antitumor effect and detailed mechanisms of Daph in ovarian cancer cells. The cytotoxic effect of Daph on ovarian cells was determined in vitro and in vivo. Cell growth, proliferation, apoptosis and ROS generation were measured by CCK8 assays, colony formation assays and flow cytometry. Western blotting was used to evaluate the related signal proteins. Immunofluorescence and transmission electron microscopy were used to evaluate markers of autophagy and autophagic flux. The antitumor effects were observed in the A2780 xenograft model. Moreover, Daph-induced autophagy was observed by enhanced LC3-II accumulation and endogenous LC3 puncta, and an autophagy inhibitor further enhanced the antitumor efficacy of Daph, which indicated that the cytoprotective role of autophagy in ovarian cancer. We found that Daph exhibited antitumor effects by inducing ROS-dependent apoptosis in ovarian cancer, which could be reversed by N-acetyl cysteine (NAC). The AMPK/Akt/mTOR pathway was involved in Daph-mediated cytoprotective autophagy, and when Daph-mediated the expression level of AMPK and autophagy were blocked, there was robust inhibition of cell proliferation and induction of apoptosis. In addition, in the A2780 xenograft model, combined treatment with Daph and an autophagy inhibitor showed obvious synergetic effects on the inhibition of cell viability and promotion of apoptosis, without any side effects. Our results suggest that Daph triggers ROS-induced cell apoptosis and induces cytoprotective autophagy by modulating the AMPK/Akt/mTOR pathway. Moreover, the combination of Daph and autophagy inhibitor may be a potential therapeutic strategy for ovarian cancer.

Artematrolide A inhibited cervical cancer cell proliferation via ROS/ERK/mTOR pathway and metabolic shift

Artematrolide A (AR-A), a guaianolide dimer isolated from Artemisia atrovirens, demonstrated significant inhibitory effect on three human hepatoma cell lines (HepG2, Huh7 and SMMC7721). The anti-cervical cancer effect and mechanism of this compound have yet to be explored. This study is to reveal the role and mechanisms of artematrolide A on cervical cancer cells, and provide the pharmacological understanding of artematrolide A. To investigate the function and possible mechanism of artematrolide A on cervical cancer cells in vitro. HeLa S3 and SiHa cells were treated with artematrolide A at various concentrations. In this study, MTT, colony formation, cell migration and invasion, cell cycle analysis, cell apoptosis, reactive oxygen species (ROS) detection, western blotting, enzyme activity, and lactate production of artematrolide A were evaluated. Artematrolide A inhibited cell viability, proliferation, migration and invasion in a dose-dependent manner, caused cell cycle arrest in G2/M phase, and induced cell apoptosis via Bcl-2/PARP-1. The mechanism of action of artematrolide A included two aspects: artematrolide A suppressed cell proliferation by activating ROS/ERK/mTOR signaling pathway and promoted glucose metabolism from aerobic glycolysis to mitochondrial respiration by activating pyruvate dehydrogenase complex (PDC) and oxoglutarate dehydrogenase complex (OGDC) via inhibiting the activity of alkaline phosphatases (ALP). Artematrolide A exhibited a significant cytotoxic activity on cervical cancer cells, induced G2/M cell cycle arrest and apoptosis by activating ROS/ERK/mTOR signaling pathway and promoting metabolic shift from aerobic glycolysis to mitochondrial respiration, which suggested artematrolide A might be a potential agent for the treatment of cervical cancer.

miRNA omics reveal neferine induces apoptosis through Ca2+mediated endoplasmic reticulum stress pathway in human endometrial cancer

Endometrial cancer (EC) as one of the most prevalent malignancies in the female reproductive system, usually has a poor diagnosis and unfavorable health effects. Neferine (Nef), derived from the edible and medicinal lotus seed, has been known for its functional activity; however, its anti-cancer mechanism for EC remains elusive. We explored the potential anti-cancer effects and underlying molecular mechanisms of Nef on EC. The cytotoxicity was tested using MTT, and the cell cycle, apoptosis, Ca Nef treatment led to Ishikawa cell apoptosis and blocked cell proliferation in the G2/M phase. In total, 101 significantly different miRNA (p 〈 0.05 and |logFC| 〉 1) were obtained and subjected to GO and KEGG enrichment analysis, which revealed the Ca Nef could block the cell cycle and induce the activation of the mitochondrial apoptotic pathway involving the Ca

Zengmian Yiliu formula suppresses cell cycle in immune-rich ovarian cancer patient-derived organoids

Ovarian cancer, often diagnosed at advanced stages, has a 5-year survival rate below 50%, indicating a critical need for innovative treatments. The Zengmian Yiliu (ZMYL) formula, a Traditional Chinese Medicine (TCM) prescription, has shown potential in enhancing chemotherapy efficacy and improving patients' quality of life, PURPOSE: To investigate the effects of the ZMYL formula on ovarian cancer organoids, focusing on its impact on organoid phenotypes and underlying mechanisms, and to explore its potential as an immunotherapeutic agent. Ovarian cancer organoids were established from surgical tissues and treated with the ZMYL formula at varying concentrations. Network pharmacology was utilized to predict the formula's therapeutic targets and pathways, and molecular docking was conducted to validate ingredient-target interactions. Phenotypic changes were monitored, and RNA sequencing was performed post-treatment to analyze gene expression alterations. A total of 34 overlapping targets of 10 compounds in the ZMYL formula and ovarian cancer were predicted by Network pharmacology analysis. The ZMYL formula induced dose-dependent morphological changes in organoids, including a reduction in size and structural sparsity at higher concentrations. RNA sequencing revealed significant modulation of cell cycle and immune response pathways, with a particular focus on immunomodulatory effects. The formula's treatment targeted key genes involved in these processes, reshaping the tumor's molecular landscape. This study establishes ZMYL's capacity to simultaneously target oncogenic drivers (e.g., cell cycle regulators) and immune checkpoints (e.g., CXCL10-mediated T cell recruitment) in ovarian cancer organoids. Unlike conventional monotherapy-focused approaches, ZMYL's multi-component mechanism offers a synergistic framework for integrating TCM with modern immunotherapies. These findings provide a foundation for future clinical evaluation of ZMYL as a precision medicine strategy to enhance treatment efficacy and mitigate chemoresistance in ovarian cancer.

Targeting and degradation of OTUB1 by Erianin for antimetastasis in esophageal squamous cell carcinoma

Berberine modulates ovarian cancer autophagy and glycolysis through the LINC01123/P65/MAPK10 signaling axis

Berberine, a readily accessible natural compound known for its ease of synthesis and low toxicity, exhibits anti-tumor properties by modulating inflammatory responses. Recent studies have revealed that berberine can also treat malignant tumors by influencing tumor metabolic reprogramming, making it a potential candidate for metabolic therapy in ovarian cancer. The anti-proliferative and anti-metastatic effects of berberine on ovarian cancer cells were investigated using CCK-8 assays, scratch assays, EDU proliferation assays, and assays related to glycolysis and autophagy. Differentially expressed lncRNAs in ovarian cancer were identified using data from the TCGA database. A specific lncRNA's role was delineated through RNA pulldown assays, silver staining, mass spectrometry analysis, CHIP assays, and immunoprecipitation experiments, focusing on its involvement in glycolysis and autophagy regulation in ovarian cancer. Additionally, the inhibitory mechanism of berberine on ovarian cancer cells was validated through cell thermal shift assays and cycloheximide protein degradation experiments to confirm its interaction with key targets. In vitro experiments revealed that berberine reduces glycolysis and autophagy levels, leading to the inhibition of ovarian cancer cell proliferation and metastasis. Bioinformatics analysis of TCGA data identified LINC00123 as associated with poor prognosis in ovarian cancer. Experimental validation, including RNA pulldown assays, confirmed that the LINC00123/P65/MAPK10 signaling axis regulates glycolysis and autophagy in ovarian cancer. Furthermore, at the molecular level, berberine inhibits the interaction between LINC00123 and P65, thereby reducing P65 protein stability and impeding its transcriptional regulation of downstream MAPK10. These findings were further validated in animal models. Our study highlights berberine's dual benefits of anti-inflammatory effects and inhibition of ovarian cancer proliferation and metastasis by modulating autophagy and glycolysis levels. Mechanistically, berberine targets the LINC00123/P65/MAPK10 signaling pathway to regulate glycolysis and autophagy in ovarian cancer. These insights not only expand the potential of berberine in ovarian cancer therapy but also provide new targets and therapeutic strategies for metabolic therapy in this cancer type.

Cucurbitacin I induces immunogenic cell death and synergistically potentiates cisplatin efficacy in epithelial ovarian cancer

Epithelial ovarian cancer (EOC) is characterized by a suppressive tumor immune microenvironment (TIME) that undermines therapeutic efficacy. Immunogenic cell death (ICD) is a promising immunotherapy strategy attributed to ameliorating the suppressive TIME. Cisplatin (CDDP), the first-line chemotherapy for EOC, is hindered by cumulative toxicity. Cucurbitacin I (CuI), a bioactive plant-derived compound, exhibits remarkable antitumor activity in preclinical models. However, its ICD-inducing capability and cisplatin-sensitizing potential in EOC remain uninvestigated. This study aims to systematically elucidate the ICD-inducing capability of CuI and its potential to enhance CDDP's antitumor effects in EOC both in vitro and in vivo. The inhibitory effects of CuI on EOC were evaluated at the cellular level. The half-maximal inhibitory concentrations (IC CuI induced caspase-3/GSDME-mediated pyroptosis in EOC cells, driving the release of ICD-related DAMPs, including surface-exposed CRT/ERp57, ATP, and HMGB1. Consequently, CuI-treated EOC cells promoted DC maturation, significantly increasing the proportion of CD80 CuI is a novel ICD inducer that elicits caspase-3/GSDME-mediated pyroptosis and boosts antitumor immunity in EOC. Notably, it enhances the chemosensitivity of EOC to CDDP, offering a promising strategy for EOC treatment.

Artesunate triggers ferroptosis in ovarian cancer via GP130-mediated IL-6/STAT3/OTUB1/SLC711 axis disruption

Ovarian cancer (OC) is highly malignant with suboptimal current therapeutic outcomes. Ferroptosis is a novel cell death mode. Artesunate (ART) has demonstrated relatively broad-spectrum anticancer effects recently, yet whether it can suppress OC by inducing ferroptosis, along with the underlying mechanisms, warrants further exploration. To clarify the anti-OC effect and mechanism of ART, focusing on ferroptosis induction and identify its therapeutic target. ART's anti-OC effects were assessed in vitro via cell counting kit-8, live-cell staining, scratch, and transwell assays. In vivo efficacy and safety of ART were evaluated in OC-bearing mice. Ferroptosis markers were analyzed by enzyme linked immunosorbent assay, western blot, and reverse transcription quantitative polymerase chain reaction, especially OTU domain-containing ubiquitin aldehyde-binding protein 1 (OTUB1)-mediated solute carrier family 7 member 11 (SLC7A11) deubiquitination was measured by co-immunoprecipitation. RNA-seq profiled transcriptional changes, and signal transducer and activator of transcription 3 (STAT3) regulation of OTUB1 was confirmed by dual-luciferase reporter and chromatin immunoprecipitation quantitative polymerase chain reaction assays. ART binding to the interleukin-6 (IL-6)/interleukin-6 receptor (IL-6R)/glycoprotein 130 (GP130) complex was determined by bioluminescence resonance energy transfer, cellular thermal shift assay, molecular docking, and dynamics simulations. GP130 point mutations validated the functional binding site. ART induced ferroptosis in OC cells by targeting GP130, inhibiting cell viability and hindering cell invasion/metastasis. In vivo, ART significantly suppressed the tumor growth of OC-bearing mice, and the tumor volume inhibitory rate and weight inhibitory rate of ART (5 mg/kg) was (82.34±4.97 %) and (78.27±4.74 %), respectively, and no side effects were observed. Mechanistically, ART bond to lysine 250 (Lys250) of GP130 (GP130 ART exerted potent anti-OC effects by targeting GP130

Chaihu Shugan San formula alleviates psychological stress-induced ovarian cancer susceptibility by inhibiting ubiquitin degradation of TLR2 in macrophages

Psychological stress is increasingly recognized for its potential to expedite tumor growth and enhance cancer susceptibility, yet the specific mechanisms at play remain largely unclear. Traditional Chinese Medicine (TCM) formulation, Chaihu Shugan San formula (CHSGS), known for its efficacy in alleviating liver Qi stagnation syndrome, has exhibited therapeutic benefits in cancer management. Macrophages are key immune cells in the tumor microenvironment, and their phagocytic and clearance functions for tumor cells play a crucial role in the occurrence and progression of tumors. In previous studies, we have found that macrophage phagocytosis is closely related to psychological stress-induced tumor susceptibility. However, the detailed comprehension of CHSGS's pharmacological mechanism and bioactive ingredients that are pertinent to its clinical use in treating ovarian cancer exacerbated by stress remains an open question in the field. This study aimed to elucidate the impact and mechanism of CHSGS in modulating psychosocial stress-associated ovarian cancer susceptibility, while also pinpointing its primary bioactive constituents responsible for mediating antitumor effects in ovarian cancer. Initially, the relationship between the macrophage phagocytosis impaired by psychological stress and the ubiquitin degradation of Toll-like receptor 2 (TLR2) using flow cytometry and co-immunoprecipitation (CO-IP). The therapeutic efficacy and mechanism of CHSGS on ovarian cancer were evaluated by the tumor volume, tumor weight, macrophage phagocytosis and CO-IP analysis. Then, the impact of the key active ingredients in CHSGS on the of TLR2/MARCH6 interaction was further elucidated through molecular docking analysis, assessment of cell viability, phagocytosis of macrophage, CO-IP analysis and microscale thermophoresis (MST) assays. Finally, the therapeutic efficacy and mechanisms of baicalin in ovarian cancer were evaluated based on the results of tumor volume, tumor weight, and macrophage phagocytosis in vivo. Our findings indicate that psychosocial stress promotes lipid peroxidation in macrophages, which in turn recruits the E3 ubiquitin ligase MARCH6 and triggers the proteasome-dependent degradation of TLR2, thereby impairing macrophage phagocytic function and increasing ovarian cancer susceptibility. CHSGS intervention effectively disrupted the binding relationship between TLR2 and MARCH6, markedly diminishing TLR2 degradation and consequently restoring the macrophage's phagocytic capability. Baicalin is a promising active ingredient of CHSGS that can alleviate stress-induced impairment of macrophage phagocytosis by inhibiting the binding of TLR2/MARCH6. Our results reveal that stress enhances susceptibility to ovarian cancer via ubiquitin degradation of TLR2 in macrophages, and indicate that the TLR2/MARCH6 complex could serve as a promising therapeutic target for CHSGS in the treating ovarian cancer.

Curcuminoids WM03 inhibits ovarian cancer cisplatin-resistant cells proliferation and reverses cisplatin resistance by targeting DYRK2

Cisplatin is a common chemotherapy agent used to treat ovarian cancer and cisplatin resistance is the most common consequence after its treatment. Curcumin has been shown to effectively inhibit the proliferation and invasion of ovarian cancer cells but its bioavailability restricts its application. The objective of this study was to develop the novel curcumin derivatives with high efficacy and synergic effects with cisplatin to inhibit cisplatin resistant ovarian cancers. Colony formation assay and growth curve assay Were used to detect cell proliferation. Transwell and cell scratch assay Were used to detect cell invasion and migration. Western blot (WB), Immunohistochemistry (IHC) and Immunofluorescence (IF) Were used to detect the expression levels of related molecules. qPCR was used to detect mRNA levels of related molecules. Kinase profile sequencing was used to analyze kinase activity. RNA seq was used to analyze significant signaling pathways. The ability of Surface plasmon resonance (SPR), Isothermal titration calorimetry (ITC) and Cellular Thermal Shift Assay (CESTA), molecular docking to analyze the binding of drugs and molecules; Co-Immunoprecipitation (Co-IP) and confocal are used to analyze intermolecular interactions. Ubiquitination is used to detect ubiquitin levels of related molecules; Animal experiments are used to simulate clinical validation RESULTS: Four curcumin derivatives Were synthesized and evaluated to treat ovarian cancers. Curcumin derivative WM03 was the most effective to inhibit A2780DR and HO8910PMDR cell proliferation with about 8-12 times more potent than curcumin. WM03 inhibited A2780DR and HO8910PMDR cell proliferation, migration, and invasion with a synergic effect of cisplatin for cisplatin resistant ovarian cells. RNA-seq results showed that the PI3K-Akt pathway differentially changed. Kinotome analysis showed that WM03 specifically targeted 4 kinases of 50 curcumin-effective kinases and dual-specificity tyrosine-(Y)-phosphorylation regulated kinase 2 (DYRK2) was the most significant kinase, The IC WM03 specifically targets DYRK2 and is more potent than curcumin to inhibit cisplatin resistant ovarian cancer cells, being a promising new drug candidate for ovarian cancers.

Dietary flavonoid intake and risk of hormone-related cancers: A population-based prospective cohort study

Dietary flavonoids may have potential effects on hormone-related cancers (HRCs) due to their anti-cancer properties via regulating hormones and suppressing inflammation and oxidative stress. We aimed to examine the association of flavonoid intake with risks of HRCs and whether this association was mediated by blood biomarkers involved in biological mechanisms. This prospective cohort study from UK Biobank included 187,350 participants free of cancer when the last dietary recall was completed. The dietary intakes of flavonoids and subclasses were assessed using 24-hour dietary recalls. Venous blood was collected at baseline and assayed for biomarkers of inflammation, hormones, and oxidative stress. Hazard ratios (HR) and 95 % confidential intervals (CI) for the associations between flavonoid intake and HRCs risk were estimated by the cause-specific Cox proportional hazards model. The role of blood biomarkers in the flavonoids-HRCs association was investigated through mediation analysis. Over a median follow-up of 9.5 years, 3,392 female breast cancer, 417 ovarian cancer, 516 endometrial cancer, 4,305 prostate cancer, 45 testicular cancer, and 146 thyroid cancer cases were documented. Compared to the lowest quintile, multivariable-adjusted HRs (95 % CIs) in the highest quintile of total flavonoid intake were 0.89 (0.80-0.99) for breast cancer, 0.68 (0.50-0.92) for ovarian cancer, and 0.88 (0.80-0.98) for female-specific cancers. For subclasses, intakes of flavonols and anthocyanidins were inversely associated with the risk of female-specific cancers (P Our findings highlighted the importance of dietary flavonoids for the prevention of HRCs in the general population, providing epidemiological evidence for dietary guidelines.

Targeting MCL1 with Sanggenon C overcomes MCL1-driven adaptive chemoresistance via dysregulation of autophagy and endoplasmic reticulum stress in cervical cancer

Cervical cancer ranks as one of the most prevalent malignancies among women worldwide and poses a significant threat to health and quality of life. MCL1 is an antiapoptotic protein closely linked to tumorigenesis, drug-resistance and poor prognosis in various cancers. Sanggenon C, a natural flavonoid derived from Morus albal., exhibits multiple activities, including anti-oxidant, anti-inflammatory, antivirus, and antitumor properties. However, the molecular mechanisms by which Sanggenon C exerts antitumor effects on in cervical cancer remain unclear. To investigate the oncogenic role of MCL1 and elucidate the antitumor activity of Sanggenon C, along with its molecular mechanisms, in cervical cancer. In vitro, the effects of Sanggenon C on proliferation, the cell cycle, apoptosis, and autophagy were explored. Transcriptome sequencing was employed to analyze critical genes and pathways. The expression of genes or proteins was evaluated via immunofluorescence, qRT-PCR, immunohistochemistry, and Western blotting. To identify targets of Sanggenon C, various techniques such as clinical database analysis, molecular docking, cellular thermal shift assays, co-immunoprecipitation, and ubiquitination assays were utilized. Additionally, Xenograft mouse models were established to further investigate Sanggenon C as a novel MCL1 inhibitor and its anti-tumor activity in vivo. Our investigation reveals that Sanggenon C effectively inhibits cervical cancer cell proliferation both in vitro and in vivo. Furthermore, Sanggenon C induces endoplasmic reticulum stress and triggers protective autophagy via activation of the ATF4-DDIT3-TRIB3-AKT-MTOR signaling axis. Furthermore, Sanggenon C specifically targets MCL1 to exert its antitumor effects by modulating MCL1 protein stability through SYVN1-mediated ubiquitination. Notably, MCL1 overexpression attenuates the Sanggenon C-induced decrease in cell viability and apoptosis. Our study further characterizes the role of MCL1 in cisplatin resistance and identifies MCL1 as a promising target for Sanggenon C, which effectively inhibits proliferation and induces apoptosis in cisplatin-resistant cervical cancer cells. Importantly, combining Sanggenon C with an autophagy inhibitor represents a promising strategy to enhance therapeutic outcomes in cisplatin-resistant cervical cancer cells. Our findings demonstrates that Sanggenon C induces endoplasmic reticulum stress and highlights the potential of targeting MCL1 to exploit vulnerabilities in drug-resistant cervical cancer cells. Sanggenon C emerges as a promising therapeutic agent against MCL1-driven adaptive chemoresistance through disruption of autophagy and endoplasmic reticulum stress in cervical cancer.

Artemisia argyi ethyl acetate fraction suppresses ovarian cancer progression via JUN/MAPK10 signaling: An integrated multi-omics investigation

Artemisia argyi, a traditional Chinese medicinal herb widely applied in gynecology, has recently attracted interest for its pharmacological activities and emerging anti-tumor potential. This study aimed to investigate the inhibitory effects and mechanisms of the ethyl acetate fraction (EA) of A. argyi on ovarian cancer (OvC). Anti-proliferative and anti-migratory effects of EA were evaluated in OvC cell lines (A2780, SKOV3) using CCK-8, colony formation, wound healing, Hoechst 33,258 staining, Annexin V-FITC/PI apoptosis assay, and Western blotting. An A2780 xenograft nude mouse model was employed to assess in vivo efficacy and safety. Integrated transcriptomic, proteomic, and metabolomic analyses were conducted to elucidate mechanisms, with key targets validated by RT-qPCR, Western blot, IHC, and IF. EA-derived components and their plasma availability were profiled using UHPLC-QE Orbitrap-MS. EA markedly inhibited OvC cell proliferation (IC₅₀ = 6.212 and 9.569 μg/ml) and migration, while inducing apoptosis and autophagy. Western blotting showed increased Bax, cleaved Caspase-3, and LC3-II, alongside reduced BCL2 expression. In vivo, EA (15/30 mg/kg) significantly suppressed xenograft tumor growth (39.47 %) without overt toxicity. Multi-omics analysis revealed regulation of Apelin, Oxytocin, cAMP signaling, and arachidonic acid metabolism, identifying JUN and MAPK10 as central targets. Downregulation of JUN, p-JUN, and MAPK10 was validated in tumor tissues by RT-qPCR, WB, IHC, and IF. Sixteen EA-derived components were detected in plasma. The EA fraction of A. argyi exerts potent anti-OvC activity by suppressing proliferation and migration, and by inducing apoptosis via JUN/MAPK10 signaling.

Indirubin suppresses ovarian cancer progression by inhibiting PI3K/AKT-mediated EMT and tumor growth without systemic toxicity

This study investigated indirubin's effects on ovarian cancer cell proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT), both in vitro and in vivo, while exploring its underlying mechanisms. The anti-tumor activity of indirubin was evaluated using ovarian cancer cell lines (A2780, SKOV-3, OVCAR-3) through MTT, colony formation, wound healing, and Transwell assays. Patient-derived organoids and a nude mouse xenograft model were employed for further validation. Mechanistic studies integrated bioinformatics analysis (SymMap database and transcriptome sequencing) with experimental assessment of EMT markers and PI3K/AKT pathway activity, including the use of specific inhibitors (MK2206, SB 216763). Indirubin significantly suppressed ovarian cancer cell proliferation, colony formation, migration, and invasion, as well as inhibited patient-derived organoid growth. In a xenograft model, indirubin effectively attenuated tumor growth without inducing pathological changes in major organs. Through SymMap database and transcriptomic analyses, indirubin was shown to modulate multiple signaling pathways, particularly impairing cell adhesion. Mechanistically, indirubin downregulated AKT and GSK-3β phosphorylation in the PI3K/AKT pathway, and upregulated E-cadherin while suppressing Vimentin and N-cadherin, thereby inhibiting EMT. Notably, indirubin, the PI3K/AKT inhibitor MK2206, and the GSK-3β inhibitor SB216763 all exhibited strong antitumor efficacy in vivo. Indirubin exerts broad anti-ovarian cancer effects by inhibiting proliferation, migration, invasion as well as EMT, with demonstrated efficacy in xenograft models and no observed organ toxicity. Its mechanistic overlap with PI3K/AKT inhibitors underscores its potential as a multitargeted therapeutic agent.

Tripterygium glycosides reverse cisplatin resistance in epithelial ovarian cancer by activating ferroptosis via two different pathways

In our previous study, tripterygium glycosides (TG) demonstrated to combine with cisplatin (DDP) in reversing DDP resistance in A2780/DDP cells by exacerbating ferroptosis via downregulation of glutathione peroxidase 4 (GPX4) and cystine/glutamate antiporter (System Xc-) expression. However, the regulation of ferroptosis-related proteins alone is insufficient to explain the reversal of DDP resistance in epithelial ovarian cancer (EOC), suggesting the involvement of additional mechanisms. This study focused on the underlying mechanisms through which TG contributes to reversing DDP resistance in EOC. In this study, using DDP-resistant A2780/DDP cells and a nude mouse xenograft model, we comprehensively investigated the underlying mechanisms by which TG reverses DDP resistance via ferroptosis regulation through combined animal and cellular research. The results demonstrated that TG can induce ferroptosis in A2780/DDP cells. Mechanistic studies confirmed that TG induces ferroptosis through a dual-pathway mechanism involving both Keap1/Nrf2/ARE and HIF-1α/ACSL4 signaling. Furthermore, TG combined with DDP co-regulates the p53/Bax/Bcl-2 pathway to enhance apoptosis, thereby strengthening the inhibitory effect of DDP on drug-resistant EOC. Animal studies confirmed that the combination of TG and DDP significantly suppressed tumor growth compared to DDP monotherapy, with no observed increase in toxicity, indicating favorable treatment safety. In conclusion, our findings reveal for the first time that the TG combined with DDP reverses EOC drug resistance through the simultaneous induction of ferroptosis (HIF-1α/ACSL4 and Keap1/Nrf2/ARE) and apoptosis (p53/Bax/Bcl-2). These results provide experimental evidence supporting the clinical application of TG in overcoming DDP chemotherapy resistance in EOC.

Herbal medicines for radiation vaginitis: A systematic review and meta-analysis

As a sequela of cervical cancer irradiation, radiation vaginitis provokes vaginal inflammation and may develop into ulcers, necrosis, adhesions and cause pain, which markedly diminishes the quality of life for affected individuals. However, the efficacy of conventional modalities, including medication and physical therapy, has not been satisfactory. Herbal medicines (HMs) demonstrate favorable safety profiles and efficacy in preventing and treating radiation vaginitis. The registration number for this systematic review is CRD42025627673. Web of Science, Scopus, Embase, PubMed, Clinicaltrials.gov, Cochrane Library, Ovid, Sinomed, PsycholNFO, Wanfang Data, ChiCTR, Chongqing VIP, CINAHL and China National Knowledge Infrastructure (CNKI) for randomised controlled trials (RCT) involving herbal products from their inception to March 1, 2025. Review Manager 5.4 and Stata 15.0 were used to assess risk of bias and conduct meta-analyses. The required information size (RIS) was calculated using TSA software (version 0.9.5.10). Subsequently, the overall certainty of evidence was evaluated with the GRADE profiler (version 3.6). 16 studies containing 18 trials with 984 patients were included. The results showed that HMs were more effective than those of the control group in treating radiation vaginitis. Key indicators of outcomes include radiation vaginitis incidence(RR = 0.51 [0.40, 0.65], p < 0.00001, I2 = 20 %) (GRADE: ⊕⊕⊝⊝low), grade 1(RR = 0.69 [0.45, 1.06], p = 0.09, I2 = 0 %) (GRADE: ⊕⊝⊝⊝very low), grade 2(RR = 0.40 [0.23, 0.67], p = 0.0005, I2 = 0 %) (GRADE: ⊕⊕⊝⊝low), grade 3(RR = 0.27 [0.10, 0.75], p = 0.01, I2 = 0 %) (GRADE: ⊕⊕⊝⊝low), grade 4 (RR = 0.33 [0.05, 2.06], p = 0.24, I2 = 0 %) (GRADE: ⊕⊝⊝⊝very low), total efficiency(RR = 1.26 [1.19, 1.32], p < 0.00001, I2 = 41 %) (GRADE: ⊕⊕⊝⊝low), recovery rate(RR = 1.58 [1.12, 2.23], p = 0.010, I2 = 75 %) (GRADE: ⊕⊝⊝⊝very low), treatment time((SMD = -6.12 [-7.94, -4.30], p < 0.00001, I2 = 85 %) (GRADE: ⊕⊝⊝⊝very low), recurrence rate(RR = 0.41 [0.26, 0.62], p < 0.0001, I2 = 0 %) (GRADE: ⊕⊝⊝⊝very low). HMs significantly reduced the incidence of radiation vaginitis in patients undergoing radiotherapy for cervical cancer, improved the severity of radiation vaginitis, increased the total efficiency, decreased the recurrence rate of treating radiation vaginitis, and further shortened the treatment time for patients with radiation vaginitis. The adverse event rate of HMs against radiation vaginitis was less than in the conventional treatment group. However, due to the quality of the literature, the study findings warrant cautious interpretation.

Melittin suppresses ovarian cancer growth by regulating SREBP1-mediated lipid metabolism

Melittin, a major peptide component of bee venom, has demonstrated promising anti-cancer activity across various preclinical cell models, making it a potential candidate for cancer therapy. However, its molecular mechanisms, particularly in ovarian cancer, remain largely unexplored. Ovarian cancer is a life-threatening gynecological malignancy with poor clinical outcomes and limited treatment options. This study evaluated the efficacy of melittin in suppressing ovarian cancer and elucidated its underlying molecular mechanisms. A subcutaneous xenograft tumor model was established using ID8 cells in C57BL/6J mice. RNA sequencing revealed that melittin's anticancer effects were associated with the downregulation of lipid metabolism, particularly fatty acid synthesis. The impact of melittin on de novo fatty acid synthesis was assessed by measuring free fatty acid (FFA), triglyceride (TG), and total cholesterol (TC) levels in ovarian cancer cells. Lipogenic gene expression and sterol regulatory element-binding protein 1 (SREBP1) were analyzed by Western blot and quantitative real-time polymerase chain reaction. The regulation of FASN transcription by SREBP1 was explored using a dual-luciferase reporter assay. Plasmid DNA transfection and the SREBP1 inhibitor Fatostatin were employed to identify the signaling pathway mediating melittin's anticancer effects. Our results confirmed that melittin significantly reduced de novo fatty acid synthesis, as evidenced by lower FFA, TG, and lipid droplet levels. Additionally, melittin inhibited the nuclear translocation of SREBP1 and specifically reduced SREBP1-mediated FASN transcription, demonstrating effects similar to those of Fatostatin. The motif (-424/-415) within the FASN promoter is a potential SREBP-1 binding site. SREBP1 overexpression through plasmid DNA transfection significantly counteracted melittin's downregulation of FASN promoter activity and counteracted its inhibitory effects on de novo fatty acid synthesis, cell proliferation, and colony formation. Our findings suggested that melittin acts as a novel modulator of the SREBP1/FASN pathway, reducing lipogenesis and inhibiting ovarian cancer growth. This study was the first to demonstrate melittin's ability to target the SREBP1/FASN axis in ovarian cancer, identifying SREBP1 as a novel therapeutic target. These results highlighted melittin as a potential therapeutic agent for ovarian cancer by attenuating SREBP1-mediated lipid metabolism and suggested novel treatment strategies for targeting ovarian cancer.

Curdione inhibits the proliferation of human uterine leiomyomas by targeting YTHDF1

Curdione, an active sesquiterpene found in Curcumae Rhizoma, exhibits strong anti-inflammatory and antitumor properties, but its mechanism of action against uterine leiomyomas (ULMs) is not yet clear. This study aims to clarify the anti-proliferative effects of curdione on ULMs and explore the molecular mechanisms, with specific focus on the role of the m6A reader protein YTHDF1. To evaluate the effects of curdione on human ULM cells, assays such as CCK-8, crystal violet staining, EdU, TUNEL, and Annexin V-FITC/PI were used to assess cell proliferation and apoptosis. Key m6A targets were identified through a comprehensive analysis involving GEO data integration, molecular docking, and surface plasmon resonance (SPR). RNA-seq and MeRIP-seq characterized N6-methyladenosine (m6A) methylation patterns and identify downstream regulatory targets. Lentiviral transfection was used to develop YTHDF1 knockdown and overexpression cell models, with target functions validated through molecular assays. The efficacy of curdione was assessed in a rat ULM model using histopathological analysis, electron microscopy, and molecular assays. Curdione exhibited an inhibitory effect on the viability of ULM cells, significantly reducing proliferation rates and inducing apoptosis. It was observed to downregulate YTHDF1 expression at both transcriptional and translational levels. MeRIP-seq analysis identified reprogramming of m6A methylation, with 231 hypermethylated and 984 hypomethylated peaks, alongside a reduction in THBS1 m6A modification and expression. Stable YTHDF1 knockdown and overexpression models were established through lentiviral transfection, confirming YTHDF1 as a pivotal mediator of curdione's anti-proliferative effects. In a rat ULM model, curdione administration resulted in a decreased uterine index, improved histopathological features, reduced collagen deposition, and normalization of serum inflammatory cytokines and sex hormones. Immunofluorescence and western blot analyses verified the co-localization and coordinated downregulation of YTHDF1 and THBS1 in uterine tissue. Curdione exerts its effects by downregulating YTHDF1, thereby influencing m6A modification and the translation of THBS1 mRNA. This study highlights curdione's potential as a therapeutic agent for ULMs and suggests that targeting YTHDF1 could be an effective management strategy.

Shikonin and cisplatin synergistically overcome cisplatin resistance of ovarian cancer by inducing ferroptosis via upregulation of HMOX1 to promote Fe2+ accumulation

Cisplatin-based chemotherapy often results in ovarian cancer (OC) chemical resistance and treatment failure. The combination of natural compounds with platinum-based agents is a new strategy for overcoming cisplatin resistance. At present, the synergistic effects and mechanism of combination of shikonin and cisplatin to overcome cisplatin resistance in OC are still unknown. This study was to evaluate the synergistic effects of shikonin and cisplatin on cisplatin-resistant OC cells and to assess the underlying molecular basis for these effects. Cell counting kit-8 assay, colony-formation assay, proteomic analysis, reactive oxygen species (ROS) detection, lipid peroxidation (LPO) detection, Fe In vitro, shikonin and cisplatin were shown to synergistically reduce the viability of cisplatin-resistant OC cells. Proteomic results demonstrated that the combination of the two drugs induced a ferroptotic process, as evidenced by increased levels of ROS, LPO, and Fe We report for the first time that the co-treatment of shikonin and cisplatin overcomes cisplatin resistance in OC through ferroptosis. Mechanistic analysis reveals the co-treatment induces ferroptosis through upregulation of HMOX1 that promotes Fe

Ginkgetin suppresses ovarian cancer growth through inhibition of JAK2/STAT3 and MAPKs signaling pathways

Ginkgo biloba L., a kind of traditional Chinese medicine, is always used to treat various diseases. Ginkgetin is an active biflavonoid isolated from leaves of Ginkgo biloba L., which exhibits diverse biological activities, including anti-tumor, anti-microbial, anti-cardiovascular and cerebrovascular diseases, and anti-inflammatory effects. However, there are few reports on the effects of ginkgetin on ovarian cancer (OC). OC is one of the most common cancers with high mortality in women. The purpose of this study was to find out how ginkgetin inhibited OC and which signal transduction pathways was involved to suppress OC. The OC cell lines, A2780, SK-OV-3 and CP70, were used for in vitro experiments. MTT assay, colony formation, apoptosis assay, scratch wound assay and cell invasion assay were used to determine the inhibitory effect of ginkgetin. BALB/c nude female mice were injected with A2780 cells subcutaneously, then treated with ginkgetin by intragastric administration. Western blot experiment was used to verify the inhibitory mechanism of OC in vitro and in vivo. We found that ginkgetin inhibited the proliferation and induced apoptosis in OC cells. In addition, ginkgetin reduced migration and invasion of OC cells. In vivo study showed that ginkgetin significantly reduced tumor volume in the xenograft mouse model. Furthermore, the anti-tumor effects of ginkgetin were associated with a down regulation of p-STAT3, p-ERK and SIRT1 both in vitro and in vivo. Our results suggest that ginkgetin exhibits anti-tumor activity in OC cells via inhibiting the JAK2/STAT3 and MAPK pathways and SIRT1 protein. Ginkgetin could be a potential candidate for the treatment of OC.

Naringenin suppresses epithelial ovarian cancer by inhibiting proliferation and modulating gut microbiota

Ovarian cancer has the highest mortality among all gynecological malignancies; currently, no effective therapeutics are available for its treatment. Naringenin has been shown to inhibit the progression of various cancers, but its inhibitory effect on ovarian cancer remains unknown. This study aimed to evaluate the inhibitory effects of naringenin on ovarian cancer and elucidate the underlying mechanisms. Cancer cell proliferation was detected by cell counting kit-8 and crystal violet assays, and the migration capability was determined by wound healing and transwell assays. Western blotting and immunohistochemistry assays were employed to determine the expression levels of the epidermal growth factor receptor, phosphatidylinositol 3-kinase (PI3K) and cyclin D1 in vitro and in vivo, respectively. An ES-2 xenograft nude mouse model was established for the in vivo experiments, and fecal samples were collected for intestinal microbiota analysis by 16S rDNA sequencing. Naringenin suppressed the proliferation and migration of A2780 and ES-2 cancer cell lines and downregulated PI3K in vitro. In animal experiments, naringenin treatment significantly decreased the tumor weight and volume, and oral administration exhibited greater effects than intraperitoneal injection. Additionally, naringenin treatment ameliorated the population composition of the microbiota in animals with ovarian cancer and significantly increased the abundances of Alistipes and Lactobacillus. Naringenin suppresses epithelial ovarian cancer by inhibiting PI3K pathway expression and ameliorating the gut microbiota, and the oral route is more effective than parenteral administration.

Matrine induces ferroptosis in cervical cancer through activation of piezo1 channel

Cervical cancer, which is a significant public health concern in women, currently lacks effective therapeutic drugs. Matrine, a constituent of the traditional Chinese herb Sophora flavescentis Radix, is known for its anti-cervical cancer properties and ability to induce programmed cell death. The induction of cancer cell ferroptosis, which is a novel cell death pattern, can become an effective clinical therapy for tumor in the future. However, the effect of matrine on ferroptosis in cervical cancer remains to be elucidated. In this study, we investigated whether matrine induces ferroptosis in cervical cancer and elucidated the underlying mechanisms. We established an SiHa-derived tumor-bearing mouse model using CB17 severe combined immunodeficient (SCID) mice and administered a group of matrine (25, 50, and 75 mg/kg) and cisplatin (2 mg/kg). We meticulously tracked alterations in body weight and tumor size and evaluated liver and kidney health using haematoxylin and eosin (H&E) staining. Using Gene Expression Omnibus (GEO) Dataset (GSE201309), we evaluated the relationship between the effects of matrine on malignant tumor cells and ferroptosis. In vitro, tetrazolium-based colorimetric (MTT), lactate dehydrogenase (LDH) and colony formation assays were used to study the effects of matrine on SiHa cell activity and cytotoxicity. We assessed ferroptosis-related protein abundance using western blotting and ferroptosis-related indices in cells using confocal immunofluorescence microscopy. The interaction of matrine with a protein linked to ferroptosis was studied using cellular thermal shift assay (CETSA). The effects of matrine on Piezo1 expression were investigated using calcium imaging. We also used Piezo1-specific siRNA to explore the role of Piezo1 in ferroptosis. Matrine administration effectively inhibited tumor growth in a SiHa-derived tumor-bearing mouse model without inducing noticeable harm. The analysis results of GEO data set show matrine-induced effects in tumor cells were indeed involved in the process of ferroptosis. Treatment with matrine resulted in a significant reduction in GPX4 protein levels and a concurrent increase in lipid peroxide and Fe Our findings indicate that matrine exerts a protective effect against cervical cancer by inducing ferroptosis through the activation of Piezo1, but not xCT or Tfr.

Asiaticoside modulates human NK cell functional fate by mediating metabolic flexibility in the tumor microenvironment

Efficacy of Chinese herbal medicines combined with high-intensity focused ultrasound for patients with uterine fibroids: A meta-analysis with core herbs exploration

Uterine fibroids (UFs) are prevalent benign tumours affecting women; they often present with symptoms such as excessive menstrual bleeding, pelvic discomfort, recurrent pregnancy loss, and infertility. High-intensity focused ultrasound (HIFU) has emerged as a non-invasive treatment option. Recent studies suggest that Chinese herbal medicines (CHMs) may enhance the efficacy of HIFU, but a comprehensive evaluation of their combined effects is lacking. To evaluate the effectiveness of CHMs combined with HIFU for UFs through a meta-analysis on randomised controlled trials (RCTs) and identify core herbal patterns that may potentiate HIFU outcomes. English and Chinese databases were searched to identify relevant studies until May 2024. The risk of bias was evaluated using the Cochrane Risk of Bias 2 tool. A meta-analysis was performed using RevMan 5.4 software. The primary outcomes were changes in UF volume and clinical efficacy; secondary outcomes were changes in uterus volume, Traditional Chinese Medicine syndrome scores, incidence of recurrences and adverse reactions. Network analysis was performed to identify core herbs. Nineteen RCTs were included. The effects of reducing UFs volumes in the CHM group were more significant than those in the control group (SMD:0.34; 95 % CI:0.44 to -0.23, p < 0.001, I² = 0 %). The clinical effectiveness rate was higher in the CHM group than with HIFU therapy alone (risk ratio: 1.13; 95 % CI: 1.08 to 1.18; p < 0.001; I This is the first meta-analysis to evaluate the clinical effectiveness of combining CHMs with HIFU for UFs, integrating both quantitative outcome and computational core herb pattern analyses to identify CHMs that may enhance the efficacy of HIFU. In addition to demonstrating superior clinical effectiveness, this study proposes a novel hypothesis: core CHMs may enhance HIFU efficacy through a dual-phase mechanism-promoting apoptosis during thermal ablation and modulating fibrotic and proliferative signaling pathways in the post-ablation phase. These findings provide an evidence-based foundation for developing integrated therapeutic strategies.

Artesunate disrupts ribosome RNA biogenesis and inhibits ovarian cancer growth by targeting FANCA

The dysregulation of ribosome biogenesis has been extensively identified in various cancers, making it emerge as a hallmark of malignant cells. This highlights the potential of targeting ribosome biogenesis as an effective approach for treating cancer patients. Although chemotherapy drugs including doxorubicin and cisplatin often target ribosome biogenesis to induce DNA damage or inhibit tumor cell proliferation, they are associated with significant side effects. This study aims to reveal the novel role of artesunate (ART), a well-known antimalarial drug, in suppressing ribosome RNA biogenesis in ovarian cancer. In this study, the inhibitory effects of ART on ovarian cancer were studied both in vitro and in vivo. The effects of ART on ribosome RNA biogenesis were detected by 5-ethynyl uridine staining, RT-qPCR, and western blotting. Drug affinity responsive target stability, mass spectrometry, molecular docking and western blotting were combined to identify ART molecular targets. Ovarian cancer cells treated with ART exhibited significant reduction in nascent rRNA synthesis, accompanied by a remarkable down-regulation of pre-rRNA and mature rRNA expression. The inhibitory effect of ART on ribosome biogenesis subsequently impaired cell proliferation, cell migration and invasion, and induced apoptosis. In eukaryotes, ribosome RNA synthesis primarily occurs in the nucleus, involving processes such as rDNA transcription, pre-rRNA splicing and the assembly of ribosome precursors with ribosomal proteins, other closely-related proteins and small nucleolar RNAs. We observed that ART inhibited the nuclear translocation of FANCA through binding to FANCA protein, consequently leading to the inhibition of ribosome RNA synthesis. Moreover, knockdown of FANCA in ovarian tumor cells resulted in reduced rRNA transcription, suppressed cell proliferation and migration, and induced apoptosis which might be mediated through the inhibition of mTOR/RPS6 activity. In vivo studies using xenograft tumors in nude mice demonstrated that ART repressed the growth of established ovarian cancer tumors. Additionally, ART treatment significantly altered FANCA protein level in these tumors, especially suppressed its nuclear localization. These findings establish ART as a potent inhibitor of ribosome biogenesis, presenting a promising therapeutic avenue for ovarian tumors with high FANCA expression or for cancer patients exhibiting abnormal activation of the mTOR-RPS6 pathway.

Enterolactone combined with m6A Reader IGF2BP3 inhibits malignant angiogenesis and disease progression in ovarian cancer

Among all gynecological cancers, ovarian cancer is the leading cause of death. Epithelial ovarian cancer (EOC) accounts for over 85 % of ovarian cancer cases and is characterized by insidious onset, early metastasis, and a high recurrence rate. Alterations in gut microbiota, often as a consequence of chemotherapy, can promote cancer development and exacerbate the disease. The m6A reader IGF2BP3 is a regulator in the occurrence and progression of various tumors and is associated with angiogenesis. Enterolactone (ENL) has demonstrated significant anti-tumor activity against various human cancers, including EOC. However, whether ENL could interact with IGF2BP3 to suppress EOC remains unclear. This study aims to investigate suppressive effects of ENL upon combining with IGF2BP3 on EOC and elucidates the underlying mechanism. The Cell Counting Kit-8 and crystal violet assays were used to assess tumor cell proliferation. Scratch and Transwell assays were employed to evaluate tumor cell migration, while tube formation assays were utilized to examine angiogenesis. Western blotting was used to measure the expression levels of IGF2BP3, VEGF, PI3K, AKT1, p-PI3K, and p-AKT1. An in vivo xenograft nude mouse model was established, fecal samples were collected, and 16S rDNA sequencing was performed to analyze gut microbiota in association with the suppressive effects of ENL and its interactions with IGF2BP3. IGF2BP3 is highly expressed in EOC and is positively correlated with poor survival in EOC patients. ENL reduces IGF2BP3 expression in EOC, thereby inhibiting the IGF2BP3-mediated VEGF/PI3K/AKT signaling pathway and suppressing the proliferation, migration, invasion, and angiogenesis of EOC. Additionally, ENL ameliorates gut microbiome, especially in conjunction with shIGF2BP3. ENL interacts with IGF2BP3 and suppresses its expression in EOC, leading to the deactivation of the IGF2BP3-mediated VEGF/PI3K/AKT signaling pathway and the subsequent inhibition of angiogenesis. The combination of ENL and shIGF2BP3 demonstrates a synergistic effect on EOC. ENL also ameliorates the gut microbiome, especially in conjunction with shIGF2BP3, to suppress EOC.

Label-free-based quantitative proteomic analysis of the inhibition of cisplatin-resistant ovarian cancer cell proliferation by cucurbitacin B

Ovarian cancer is a serious threat to women's health, and resistance to chemotherapeutic drugs constitutes one of the principal reasons for ovarian cancer recurrence and the low overall survival rate. Therefore, it is of paramount importance to develop additional and more-effective drugs to combat resistance to chemotherapeutic drugs. Cucurbitacin B (CuB) is a natural compound found in food plants such as bitter gourd and pumpkin, and it manifests favorable antitumor effects on a variety of malignant tumors. The present study aimed to determine the mechanism effects of CuB overcomes tumor-drug resistance in ovarian cancer. We used CCK-8, Edu, flow cytometric assays and cisplatin-resistant ovarian cancer xenograft mouse model to evaluate the cellular proliferation, cellular apoptosis.and tumor growth. We subsequently applied a pharmacoproteomic approach to analyze the molecular mechanisms by which CuB inhibited the proliferation of cisplatin-resistant ovarian cancer cells. We also employed western blot and molecular docking experiments to verify elements of PI3K/Akt/mTOR pathway expression. We found that CuB inhibited cellular proliferation and promoted apoptosis in cisplatin-resistant ovarian cancer cell lines. We discerned that CuB inhibited tumor growth of xenograft mouse tumors. We ascertained that treatment of A2780-DDP cells with CuB resulted in the differential expression of 305 proteins, with 202 proteins downregulated and 103 proteins upregulated. Of these proteins, the mTOR protein was significantly downregulated in the drug-treated group. We also found that CuB inhibited PI3K, Akt, and mTOR and that it activated cGAS expression upstream of PI3K and inhibited ATR expression. Molecular docking experiments revealed that CuB was hydrogen-bonded to mTOR proteins at Gly (2142) and Thr (2207), with a binding force of -10.2 kcal/mol. Our study confirmed that cucurbitacin B inhibits the PI3K/Akt/mTOR signaling pathway, targets mTOR, suppresses the proliferation of cisplatin-resistant ovarian cancer cells.And we also found that cucurbitacin B induces DNA damage, activates cGASA and recruits IKBα,playing a crucial role in eliciting anti-tumor immunity. We herein uncovered a new use for CuB in inhibiting tumor-drug resistance, providing a novel approach to overcoming chemotherapeutic drug resistance in ovarian cancer.

Toosendanin reduces cisplatin resistance in ovarian cancer through modulating the miR-195/ERK/β-catenin pathway

Cisplatin (DDP) resistance is prevalent in ovarian cancer (OC) patients and contributes to the poor prognosis. Therefore, it is of great significance to develop new agent to intervene and even reverse DDP resistance in OC. Toosendanin (TSN), a triterpenoid extracted from the bark or fruits of Melia toosendan Sieb et Zucc, has been proved to possess significant antitumor activities. However, the efficacy of TSN on DDP resistance in OC has not been reported yet. The aim of this study is to investigate the effects of TSN on DDP resistance in OC and explore the molecular mechanism in vitro and in vivo. Human OC cell line (SKOV3) and DDP-resistant cell line (SKOV3/DDP) were used. Cell proliferation was measured by CCK-8 and colony formation assay. Annexin V/PI double staining and hoechst 33342 nuclear staining were employed to detect cell apoptosis. Transwell and wound-healing assay were used to determine the invasion and migration potential of cells respectively. Quantitative real-time PCR (qPCR) and western blotting were performed to detect the expression of molecules related to miR-195/ERK/β-catenin pathway. The effects and mechanism of TSN on DDP resistance of OC in vivo was investigated using xenograft model, TUNEL staining assay and immunohistochemistry. TSN improved the DDP sensitivity of SKOV3/DDP cells in vitro and in vivo, reflected in promoting inhibition of proliferation, invasion, migration and epithelial mesenchymal transformation (EMT) as well as induction of apoptosis by DDP. TSN could modulate the miR-195/ERK/β-catenin axis by upregulating the miR-195-5p expression and then suppressing ERK/GSK3β/β-catenin pathway which were activated in SKOV3/DDP cells. Moreover, co-treatment of β-catenin pathway activator LiCl or miR-195-5p silencing partially recovered the DDP resistance which was previously repressed by TSN. Both in vitro and in vivo data demonstrated that TSN could reduce DDP resistance in OC through regulating the miR-195/ERK/β-catenin pathway, highlighting the potential of TSN as an effective agent for favoring overcoming clinical DDP resistance in OC.

Agrimonolide inhibits cancer progression and induces ferroptosis and apoptosis by targeting SCD1 in ovarian cancer cells

Ovarian cancer is a gynaecological tumour has high incidence and mortality rates. Agrimonolide, isolated from Agrimonia pilosa Ledeb, has multiple biomedical activities, including anticancer activity. Here, we aimed to reveal the function of agrimonolide on ovarian cancer progression. MTT assay, colony-formation assay, flow cytometry, transwell assay, scratch test, western immunoblotting, reactive oxygen species (ROS) detection, and ferroptosis analysis were performed to reveal the role and underlying mechanisms of agrimonolide in ovarian cancer cell lines (A2780 and SKOV-3). The effects of agrimonolide on the SKOV-3 xenograft model were also studied. Agrimonolide dose-dependently inhibited proliferation, migration, and invasion and promoted apoptosis in A2780 and SKOV-3 cells. Agrimonolide induced ferroptosis in tumour cells, evidenced by the increased levels of ROS, total iron, and Fe Our study is the first to suggest that agrimonolide acts as a novel apoptosis- and ferroptosis-inducing agent in ovarian cancer cells by targeting SCD1. Agrimonolide may be a novel therapeutic agent for treating ovarian cancer.

Damnacanthal isolated from morinda species inhibited ovarian cancer cell proliferation and migration through activating autophagy

Ovarian cancer is a very common gynecological malignant tumor. Natural products are important sources of chemotherapy drugs for ovarian cancer. Damnacanthal is an anthraquinone derivative with anti-cancer pharmacological properties. This study aimed to investigate the mechanisms underlying damnacanthal's effects against ovarian cancer. In vitro experiments, CCK8, colony formation and flow cytometry assays were used to evaluate the anti-ovarian cancer effect of damnacanthal on SKVO3 and A2780 cells. The wound healing tests and the transwell invasion assays were used to detect the migration and infiltration of ovarian cancer cells. Western Blot assays and immunofluorescence staining were used to measure autophagy levels. In vivo experiments, the anti-ovarian cancer effect of damnacanthal was further evaluated in a xenograft nude mouse model of SKVO3 cells. Damnacanthal induced significant cell death and apoptosis, as well as significant inhibition in migration and invasion, in SKVO3 and A2780 cells, Furthermore, damnacanthal induced cell cycle arrest by increasing the protein levels of p27Kip1 and decreasing cyclin D1 levels. In addition, damnacanthal induced a significant accumulation of autophagosomes, accompanied with an increase in LC3II protein levels, and a decrease in p62 protein levels. 3-methyladenine, an autophagy formation inhibitor, significantly mitigated the damnacanthal-induced apoptosis and migration hindrance, as well as the decline in cell viability. Furthermore, the inactivation of ERK and its downstream effector mTOR signaling pathways, rather than Akt or P38 pathway, were involved in damnacanthal's activation in autophagy. In addition, TBHQ, an ERK activator, significantly inhibited damnacanthal-boosted LC3 II levels and autophagosome accumulation, and reversed damnacanthal-induced cell death, apoptosis, cell cycle arrest and migration hindrance. Finally, the anti-ovarian cancer effect of damnacanthal was confirmed in the orthotopic xenograft model of SKVO3 cells in nude mice, with tumor growth being significantly inhibited comparably to the efficacy of cisplatin. Damnacanthal was also synergistic with cisplatin and showed inhibition in cisplatin-resistant ovarian cancer cells. Damnacanthal inhibited the growth of ovarian cancer via the ERK/mTOR/autophagy signaling cascade, indicating that it may be a potential anti-ovarian cancer drug candidate.

Icaritin inhibits endometrial carcinoma cells by suppressing O-GlcNAcylation of FOXC1

Icaritin has a wide range of pharmacological activities, including significant an-titumor activity. However, the mechanism of action of icaritin in endometrial cancer (UCEC) remains unknown. FOX proteins are a highly conserved transcription factor superfamily that play important roles in epithelial cell differentiation, tumor metastasis, angiogenesis, and cell cycle regulation. FOXC1 is an important member of the FOX protein family. FOXC1 is aberrantly expressed in endometrial cancer and may play a role in the migration and invasion of endometrial cancer; however, its mechanism of action has not yet been reported. O-GlcNAc glycosylation is a common post-translational modification. In endometrial cancer, high levels of O-GlcNAcylation promote cell proliferation, migration, and invasion. Cancer development is often accompanied by O-GlcNAc modification of proteins; however, O-GlcNAc modification of the transcription factor FOXC1 has not been reported to date. To investigate the inhibitory effects of icaritin on RL95-2 and Ishikawa endometrial cancer cells in vitro and in vivo and to elucidate the possible molecular mechanisms. CCK8, colony formation, migration, and invasion assays were used to determine the inhibitory effects of icaritin on endometrial cancer cells in vitro. Cell cycle regulation was assayed by flow cytometry. Protein levels were measured based on western blotting. The level of FOXC1 expression in endometrial cancer tissues was determined by immunohistochemistry. To assess whether icaritin also has activity in vivo, its effect on tumor xenografts was evaluated. Immunohistochemical analysis of clinical samples revealed that FOXC1 expression was significantly higher in endometrial cancer tissues than in normal tissues. Downregulation of FOXC1 inhibited the proliferative, colony formation, migration, and invasive abilities of RL95-2 and Ishikawa endometrial cancer cells. Icaritin inhibited the proliferation, colony formation, migration, and invasion of endometrial cancer cells and blocked the cell cycle in S phase. Icaritin affected O-GlcNAc modification of FOXC1 and thus the stability of FOXC1, which subsequently triggered the inhibition of endometrial cancer cell proliferation. The anti-endometrial cancer effect of icaritin is related to the inhibition of abnormal O-GlcNAc modification of FOXC1, which may provide an important theoretical foundation for the use of icaritin against endometrial cancer.