European Journal of Pharmacology

TGFαL3-SEB fusion protein as an anticancer against ovarian cancer

TGFαL3-SEB is a new synthetic fusion protein produced by the combination of the third loop of transforming growth factor with staphylococcal enterotoxin type B. In the current study, the anti-tumor effects of TGFαL3-SEB were evaluated against SKOV3 cells, which highly expressed the epidermal growth factor receptor (EGFR). Our findings showed that incubation of SKOV3 cells with 75, 100 and 150 μg/ml of TGFαL3-SEB significantly reduces the proliferation rate in a concentration-dependent manner (P < 0.05) and its IC50 value was 110 μg/ml. Caspase-3 activity was increased from 100% for control cells to 109, 144, and 169% for 75, 100 and 150 μg/ml of TGFαL3-SEB treatment, respectively. Caspase-9 activity and bax/bcl-2 ratio were also confirmed the apoptosis induction ability of TGFαL3-SEB (P < 0.001). Flow cytometry examination also showed that apoptosis was induced and the number of apoptotic cells was increased from 8.2% in un-treated cells to 20.9, 50, and 90% in response to 75, 100 and 150 μg/ml of TGFαL3-SEB fusion protein in a concentration-dependent manner (P < 0.05). The mRNA expression level of VEGF was also reduced to 0.89, 0.69, and 0.60, respectively in response to 75, 100 and 150 μg/ml of TGFαL3-SEB fusion protein exposure, respectively (P < 0.5). In summary, the findings of our study uncovered that TGFαL3-SEB fusion protein induced apoptosis and reduced angiogenesis in SKOV3 ovarian cancer cells in a concentration-dependent manner. This protein has the potential to act against EGFR expressing malignant cells to serve as a pro-apoptotic and angiogenesis blocker agents; however, further studies are needed to confirm its ability.

Small interfering RNA (siRNA) based therapies for cancer treatment: A special focus on cervical cancer

Small interfering RNA (siRNA)-based therapies are rapidly advancing as a novel approach in cancer treatment by selectively silencing oncogenes and disease-associated genes via the RNA interference (RNAi) pathway. By harnessing the body's natural cellular machinery to degrade specific messenger RNA (mRNAs), siRNA therapies prevent the production of harmful proteins with remarkable precision, offering reduced off-target effects compared to conventional chemotherapy or radiation. This review emphasizes the growing importance of siRNA in various cancers, particularly cervical cancer, which is often linked to persistent Human Papillomavirus (HPV) infections. It discusses how siRNA therapies target key viral oncogenes like E6 and E7, reactivating tumour suppressor pathways, modulating immune responses and improving sensitivity to chemotherapy. In addition, the review explores recent advancements in siRNA delivery strategies such as lipid nanoparticles, polymeric carriers, dendrimers and exosomes that address major challenges like nuclease degradation, limited cellular uptake and immune activation. The clinical potential of siRNA, including its integration with personalized medicine and ongoing clinical trials, is evaluated, alongside a critical analysis of current limitations such as immunogenicity, delivery efficiency and manufacturing complexities. Altogether, the review highlights both the promises and challenges of siRNA-based therapies and points towards future directions needed to realize their full clinical potential in oncology.

C1632 inhibits ovarian cancer cell growth and migration by inhibiting LIN28 B/let-7/FAK signaling pathway and FAK phosphorylation

The highly conserved RNA-binding protein LIN28B and focal adhesion kinase (FAK) are significantly upregulated in ovarian cancer (OC), serving as markers for disease progression and prognosis. Nonetheless, the correlation between LIN28B and FAK, as well as the pharmacological effects of the LIN28 inhibitor C1632, in OC cells have not been elucidated. The present study demonstrates that C1632 significantly reduced the rate of DNA replication, arrested the cell cycle at the G0/G1 phase, consequently reducing cell viability, and impeding clone formation. Moreover, treatment with C1632 decreased cell-matrix adhesion, as well as inhibited cell migration and invasion. Further mechanistic studies revealed that C1632 inhibited the OC cell proliferation and migration by concurrently inhibiting LIN28 B/let-7/FAK signaling pathway and FAK phosphorylation. Furthermore, C1632 exhibited an obvious inhibitory effect on OC cell xenograft tumors in mice. Altogether, these findings identified that LIN28 B/let-7/FAK is a valuable target in OC and C1632 is a promising onco-therapeutic agent for OC treatment.

IL-6 promotes nuclear translocation of HIF-1α to aggravate chemoresistance of ovarian cancer cells

The inflammatory milieu in tumor modulates the resistance to the conventional antitumoral therapies. Interleukin-6 (IL-6), a pleiotropic pro-inflammatory cytokine and a crucial mediator of tumor development, has been targeted as a therapeutic strategy to overcome chemoresistance in the treatment of tumors. The protein levels and nuclear translocation of HIFs (hypoxia-inducible factors), such as HIF-1α, are linked to the drug resistance of tumor cells. However, whether IL-6 promotes the nuclear translocation of HIF-1α and the related mechanism remain to be investigated. We applied two ovarian cancer (OvCa) cell lines, A2780 cells and SKOV3 cells for the in vivo and in vitro studies. We found that IL-6 up-regulates the HIF-1α expression via the signal transducer and activator of transcription 3 (STAT3) signaling under hypoxia in either endogenous or exogenous way, and then we proved that IL-6 enhances the transcriptional activity of HIF-1α via the STAT3 signaling. Further mechanism research revealed that IL-6 promotes the nuclear translocation of HIF-1α through the STAT3 signaling under hypoxia. Proliferation assay and apoptosis assay were applied and proved that IL-6 enhances the chemoresistance of OvCa cells against cisplatin through the upregulation of HIF-1α via the STAT3 signaling in vitro. The In vivo studies confirmed the effect of IL-6 in increasing the chemoresistance of OvCa cells against cisplatin through the IL-6/STAT3/HIF-1α loop in the animal models. Our data elucidates the explicit mechanism of IL-6/STAT3/HIF-1α loop in OvCa and also provides new insights into the development of different approaches for the inflammation-induced and hypoxia-induced resistance in tumor therapies.

Confirming whether novel rhein derivative 4a induces paraptosis-like cell death by endoplasmic reticulum stress in ovarian cancer cells

Ovarian cancer is the leading cause of death among gynecologic cancer patients. Although platinum-based chemotherapy as a frontline treatment for ovarian cancer has been widely used in clinical settings, its clinical efficacy is not satisfactory due to the resistance of ovarian cancer cells to apoptosis. Therefore, it is of great significance to induce non-apoptotic programed cell death patterns, such as paraptosis, in ovarian cancer. In this study, we aimed to explore the potential anticancer mechanisms of novel rhein derivative 4a, which was modified with rhein as a lead compound. The results showed that a wide range of vacuoles from the endoplasmic reticulum and mitochondria appeared in ovarian SKOV3, SKOV3-PM4, and A2780 cells treated with derivative 4a, and the cell death caused by derivative 4a is a type of non-apoptotic and non-autophagic death, which is caused by expansion and damage of the endoplasmic reticulum or mitochondria, showing the characteristics of para-apoptotic death. Furthermore, derivative 4a stimulated the unfolded protein reaction of ovarian cancer cells by upregulating the expression of Bip78 and activating the PERK-eIF2α-ATF4 pathways. Notably, rhein derivative 4a-induced cell death was positively correlated with activation of p38, ERK, and JNK, and negatively correlated with Alix, a known protein that inhibits paraptosis. In addition, derivative 4a treatment also induced G2/M phase arrest in ovarian cancer cells. Taken together, our study reveals that derivative 4a induces paraptosis, and this finding can serve as a basis in developing a new strategy for the treatment of antiapoptotic ovarian cancer.

Osthole inhibits ovarian carcinoma cells through LC3-mediated autophagy and GSDME-dependent pyroptosis except for apoptosis

Ovarian carcinoma (OC) begins in the ovaries and remains a highly lethal malignancy. Despite great efforts have been made to fight against OC, there still remain limited therapeutic options owing to chemotherapy drug resistance and serious side effects. Osthole is a derivative of coumarin and extracted from Cnidium monnieri (L.) Cusson, which has been drawn more attention due to its high biological activity in various disease. However, the underlying mechanism of osthole in OC is still unclear. In this study, we aim to evaluate the mechanism of osthole against OC cells. Methodologically, Cell Counting Kit-8 (CCK-8) and LIVE/DEAD™ Cell Imaging experiments were employed to assess cell viability. 2',7'-Dichlorofluorescin diacetate (DCFH-DA) staining, flow cytometry, Hoechst staining, JC-1 staining assay and western blotting were performed to study apoptosis. Transmission electron microscopy, western blotting and monodansyl cadaverine (MDC) staining assay were used to study autophagy. Western blotting and microscopy image were employed to determine pyroptosis. Our results demonstrated that osthole could significantly suppress OC cells growth in a dose-dependent manner. We further proved that osthole could inhibit OC cells growth by mitochondria-mediated apoptosis. Meanwhile, we also discovered that osthole could trigger cell autophagy and lead to cell death. Furthermore, our study revealed that osthole could lead to pyroptosis through inducing the cleavage of gasdermin E (c-GSDME) level. Taken together, Osthole could significantly suppress the growth of OC cells and induce OC cells death via apoptosis, pyroptosis and autophagy, which is a promising new drug for the treatment of OC.

Glucosamine reverses drug resistance in MRP2 overexpressing ovarian cancer cells

Glucosamine (GlcN), a natural amino sugar in human body, was reported to exhibit anticancer activity against some tumors. In the present study, we evaluated the cytotoxicity and multi-drug resistance (MDR) reversal activity of GlcN on resistant MRP2-overexpressing ovarian cancer A2780RCIS cells. The cytotoxicity and MDR reversal activity of GlcN on cancer cells were measured by MTT assay. The effects of GlcN on MRP1 and MRP2 mRNA expression and function were evaluated by qRT-PCR and flow cytometry, respectively. The cell migration capacity of ovarian cancer cells were assessed in the presence or absence of GlcN using wound healing migration assay. Furthermore, the effects of GlcN on the mRNA expression of E-cadherin, vimentin and α-smooth muscle actin as Epithelial-Mesenchymal Transition (EMT)-related markers were evaluated by qRT-PCR. Our results indicated that glucosamine reduced the proliferation of human ovarian cancer cell lines (A2780) and its cisplatin resistant variant (A2780RCIS) in a dose-dependent manner. The IC50 values for A2780RCIS cells treated with cisplatin in the presence of different concentrations of GlcN (0, 1, 2 and 3 mM) for 72 h were 44.463 ± 1.603, 35.17 ± 0.025, 22.25 ± 0.018, 17.78 ± 0.012 μM respectively. Also GlcN decreased the expression of MRP1 and MRP2 mRNA in ovarian cancer cells. Our results further demonstrated that although GlcN had no significant effects on the expression of studied EMT-related markers in invasive A2780RCIS cells, it was able to inhibit their migration in vitro. According to these findings, GlcN could effectively enhance cisplatin cytotoxicity in resistant A2780RCIS cells.

Ferrocenes as new anticancer drug candidates: Determination of the mechanism of action

Chemotherapy plays an essential role in the management of cancer worldwide. However, it is a non-specific treatment limited by major drawbacks, thus identification and testing of new promising molecular structures representing potential drug candidates are urgently needed. In this work, ferrocene complexes as potential antitumor drugs that display cytotoxicity in low micromolar concentrations against ovarian cancer cells A2780 and SK-OV-3 were investigated to identify their mode of action. Their mechanism of cellular accumulation was studied using differential pulse voltammetry and inductively coupled plasma - mass spectrometry. Their mode of cell death induction was determined by changes in the mitochondrial membrane potential, production of reactive oxygen species and by Annexin V staining. Transferrin receptors were identified as key mediators of intracellular accumulation of ferrocenes and the extent of cellular uptake reflected the anticancer activity of individual compounds. Functional analysis revealed activation of intrinsic apoptosis as a dominant mechanism leading to regulated cell death induced in ovarian cancer cells by ferrocenes. Ferrocenes represent a group of promising sandwich organometallic complexes exerting cytotoxic activity. We suggest their application not only as standalone chemotherapeutics but also as modifying substituents of known drugs to improve their antitumor effects.

Statins as adjuvants in the treatment of ovarian cancer: Controversy and misunderstanding

Ovarian cancer is frequently detected in advanced stages when the chances of survival are very low. Although chemotherapy is the treatment of choice, it is often rapidly compromised by the development of chemoresistance in patients. There are few pharmacological alternatives for managing chemoresistant ovarian cancer and statins have been suggested as an alternative, but their use is considered controversial. We present an overview of the most relevant epidemiological, in vitro and in vivo studies on the effects of statins in mono- or polytherapy for ovarian cancer. We conclude that the negative or inconclusive results of some epidemiological studies on statin-based cancer treatment are probably due, in large part, to the low doses given to patients, equivalent to those prescribed for hypercholesterolemia. Higher concentrations are well tolerated in animal models and by most patients in clinical trials. Future research is necessary to explore this possibility.

Prospects of GPR68 in combined chemo-immunotherapy of tumors

Potential anti-ovarian cancer agent: Isoaaptamine interference epithelial-mesenchymal transition, induces intrinsic apoptosis, and suppresses angiogenesis

Ovarian cancer (OC) often metastasizes to the peritoneal cavity, causing late-stage diagnosis with high mortality. Aaptamine and isoaaptamine, sponge-derived isomeric alkaloids, were investigated for the anti-tumor potential of isoaaptamine in OC cells. Isoaaptamine outperforms aaptamine, significantly reducing the viability of PA-1 and SKOV-3 cells with half maximal inhibitory concentration (IC

Luteolin modulates the TGFB1/PI3K/PTEN axis in hormone-induced uterine leiomyomas: Insights from a rat model

Uterine leiomyomas (UL), or fibroids, are non-cancerous tumors of the uterine smooth muscle, affecting approximately 70% of women of reproductive age. They are the most prevalent solid tumors in the gynecological tract and a major indication for hysterectomy. The pathogenesis of UL involves uterine inflammation, uncontrolled cell division, and suppressed apoptosis. This study evaluated the protective effects of luteolin, a flavonoid known for its anti-inflammatory and antioxidant properties, against diethylstilbestrol and progesterone-induced UL in female rats. Twenty-four female Wistar rats were divided into four groups: (1) control, (2) luteolin (10 mg/kg, PO), (3) UL (diethylstilbestrol 1.35 mg/kg + progesterone 1 mg/kg, SC), and (4) UL + luteolin (10 mg/kg). The treatment duration was five weeks. Histological analyses were performed using hematoxylin and eosin (H&E) staining and Masson's Trichrome staining to evaluate uterine architecture and fibrosis. Histological results demonstrated normal uterine architecture in the control and luteolin groups, with marked neoplastic cell proliferation and fibrosis in the UL group, significantly mitigated by luteolin treatment. Luteolin reduced uterine weights and exhibited antioxidant, anti-inflammatory, pro-apoptotic, and anti-proliferative effects. Immunohistochemical analysis revealed that luteolin significantly reduced α-SMA protein expression, suggesting its role in modulating fibrotic pathways by inhibiting TGF-β1 and PI3K and enhancing PTEN production. These findings highlight luteolin's potential as a non-invasive therapeutic option for UL and suggest the need for further clinical studies to establish its efficacy, optimize dosage, and evaluate its safety profile in humans.

Berberine inhibits chemotherapy-exacerbated ovarian cancer stem cell-like characteristics and metastasis through GLI1

Despite the remarkable clinical response in ovarian cancer therapy, the distinctively high metastasis rate is still a barrier to achieve satisfying prognosis. Our study aimed to decipher the role of berberine in inhibiting chemotherapy-exacerbated ovarian cancer metastasis. We found that chemotherapy exacerbated the migration and cancer stem cell (CSC)-like characteristics through transcriptional factor GLI1, which regulated the pluripotency-associated gene BMI1 and the epithelial-mesenchymal transition (EMT) markers Vimentin and Snail. Berberine could not only down-regulate CSC-like characteristics but also reverse EMT and migration through inhibiting chemotherapy-activated GLI1/BMI1 signaling pathway. Together, our study revealed the pivotal role of berberine in overcoming chemotherapy-exacerbated ovarian cancer metastasis, thereby provided a potential adjuvant therapeutic agent in combination with chemotherapeutics to prevent metastasis during ovarian cancer chemotherapy.

Selective therapeutic benefit of X-rays and inhibitors of EGFR, PI3K/mTOR, and Bcl-2 in breast, lung, and cervical cancer cells

Cancer continues to be a growing burden, especially in the resource limited regions of the world, and more effective and affordable therapies are highly desirable. In this study, the effect of X-ray irradiation and four inhibitors, viz. those against epidermal growth factor receptor (EGFR), phosphatidylinositol 3-kinase (PI3K), mammalian target of rapamycin (mTOR) and B-cell lymphoma 2 (Bcl-2) was evaluated in lung, breast, and cervical cancer cell lines, including normal cell lines to determine and compare the potential therapeutic benefit of these treatment modalities. A clonogenic survival assay was used to determine the radiosensitivity and cytotoxicity of inhibitors of EGFR, PI3K/mTOR, and Bcl-2 in the cell lines. From the data, the equivalent dose at which 50% of the cell populations were killed, for cancer and normal cells, was used to determine the relative cellular sensitivity to X-ray irradiation and inhibitor treatment. It was found that breast cancer cell lines were more sensitive to X-ray irradiation, whilst cervical and lung cancer cell lines were more sensitive to EGFR and PI3K/mTOR inhibitor therapy. These data suggest that patients with breast cancer possessing similar characteristics to MDA-MB-231 and MCF-7 cells may derive therapeutic benefit from X-ray irradiation, whilst EGFR and PI3K/mTOR inhibitor therapy may potentially benefit cancer patients possessing cancers similar to HeLa and A549 cells.

RETRA induces necroptosis in cervical cancer cells through RIPK1, RIPK3, MLKL and increased ROS production

Cervical cancer is the fourth most prevalent cancer in women worldwide, predominantly infected with human papillomavirus (HPV). The current chemo and radiotherapies are mostly futile due to acquired resistance to apoptosis and warrant new therapeutic approaches targeting potent non-apoptotic cell death pathways to eliminate cervical cancer cells. Induction of necroptosis by pharmaceutical interventions is emerging as a promising tool in multiple apoptotic resistant cancer cells. RETRA (REactivation of Transcriptional Reporter Activity) is a small molecule known to induce expression of p53 regulated genes in mutant (mt) p53 cells but, detailed mechanisms of its anticancer effects are poorly known. The present study investigated the potentials of RETRA as an anticancer agent and found that it induces necroptosis selectively in cervical cancer cells irrespective of p53 status through the phosphorylation of receptor-interacting protein kinase 1,3 (RIPK1, RIPK3) and mixed lineage kinase domain-like protein (MLKL) with no cytotoxic effects in normal human peripheral blood mononuclear cells (PBMCs). RETRA-treated cells also displayed necroptotic morphology of disintegrated plasma membranes with intact nuclei and also showed cell cycle arrest at the S phase with the upregulation of p21 and downregulation of cyclin-D3. Intriguingly, the combinatorial approach of using RETRA with Necrostain-1, a known inhibitor of necroptosis, reversed the effect of RETRA and rescued cell death. Moreover, induction of necroptosis by RETRA is associated with mitochondrial hyperpolarization and elevated ROS production. Collectively, these findings suggest that RETRA induces cell death via necroptosis with increased production of ROS, accentuating the therapeutic implication of RETRA in cervical cancer cells.

Targeted drug delivery in cervical cancer: Current perspectives

Cervical cancer is preventable yet one of the most prevalent cancers among women around the globe. Though regular screening has resulted in the decline in incidence, the disease claims a high number of lives every year, especially in the developing countries. Owing to rather aggressive and non-specific nature of the conventional chemotherapeutics, there is a growing need for newer treatment modalities. The advent of nanotechnology has assisted in this through the use of nanocarriers for targeted drug delivery. A number of nanocarriers are continuously being developed and studied for their application in drug delivery. The present review summarises the different drug delivery approaches and nanocarriers that can be useful, their advantages and limitation.

BNIP3 as a potential target of esculetin for treating ovarian cancer and prognostic biomarker in ovarian cancer patients

Among gynecological tumors, ovarian cancer has the highest mortality rate and worst prognosis. Therefore, it is crucial to identify and develop novel methods and targets for treating ovarian cancer. Previous studies have shown that esculetin exerts antitumor effects in a variety of cancers, however, its anti-ovarian cancer effects and mechanisms of action remain unclear. In the present study, we investigated the anti-ovarian cancer effects and mechanisms of esculetin in A2780 and OVCAR3 ovarian cancer cells and established an xenograft ovarian cancer mouse model. Esculetin significantly inhibited ovarian cancer cell proliferation, blocked cell cycle progression, and promoted apoptosis and DNA damage in a concentration-dependent manner. Furthermore, esculetin inhibited tumor growth in an xenograft ovarian cancer mouse model. Moreover, RNA-seq showed that 2114 genes were significantly altered in A2780 cells after esculetin treatment. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis indicated that these differentially expressed genes were mainly enriched in the calcium, HIF-1, and Rap1 signaling pathways. Interestingly, BNIP3 expression was notably upregulated in ovarian cancer cells after esculetin treatment. Finally, we found that low BNIP3 expression was correlated with poor prognosis in patients with ovarian cancer. These results prove that esculetin may be a valuable anti-ovarian cancer drug, and that BNIP3 is a potential treatment target for esculetin and a potential prognostic biomarker for ovarian cancer.

Pixantrone as a novel MCM2 inhibitor for ovarian cancer treatment

Mini-chromosome maintenance protein 2 (MCM2) is a potential target for the development of cancer therapeutics. However, small molecule inhibitors targeting MCM2 need further investigation. Molecular dynamics simulation was performed to identify active pockets in the MCM2 protein structure (6EYC). The active pocket was used as a docking model to discover MCM2 inhibitors by using structure-based virtual screening and surface plasmon resonance (SPR) assay. Furthermore, the efficacy of pixantrone targeting MCM2 in ovarian cancer was evaluated in vitro and in vivo. Pixantrone was identified as a novel inhibitor of MCM2 by virtual screening. SPR binding affinity analysis confirmed the direct binding of pixantrone to MCM2 protein. Pixantrone significantly reduced the viability of ovarian cancer cells A2780 and SKOV3 in a dose- and time-dependent manner. In addition, pixantrone inhibited DNA replication, and induced cell cycle arrest and apoptosis in ovarian cancer cells via targeting MCM2. Knockdown of MCM2 could attenuate the inhibitory activity of pixantrone in ovarian cancer cells. Furthermore, pixantrone significantly suppressed ovarian cancer growth in the A2780 cell xenograft mouse model and showed favorable safety. These findings suggest that pixantrone may be a promising drug for ovarian cancer patients by targeting MCM2 in the clinic.

Targeting the NRF2/KEAP1 pathway in cervical and endometrial cancers

Cervical and endometrial cancers are among the most dangerous gynaecological malignancies, with high fatality and recurrence rates due to frequent diagnosis at an advanced stage and chemoresistance onset. The NRF2/KEAP1 signalling pathway plays an important role in protecting cells against oxidative damage due to increased reactive oxygen species (ROS) levels. NRF2, activated by ROS, induces the expression of antioxidant enzymes such as heme oxygenase, catalase, glutathione peroxidase and superoxide dismutase which neutralize ROS, protecting cells against oxidative stress damage. However, activation of NRF2/KEAP1 signalling in cancer cells results in chemoresistance, inactivating drug-mediated oxidative stress and protecting cancer cells from drug-induced cell death. We review the literature on the role of the NRF2/KEAP1 pathway in cervical and endometrial cancers, with a focus on the expression of its components and downstream genes. We also examine the role of the NRF2/KEAP1 pathway in chemotherapy resistance and how this pathway can be modulated by natural and synthetic modulators.

The role of bromodomain-containing protein 4 in the replication human papillomavirus, apoptosis, proliferation and migration of cervical cancer cells

We investigated the correlation between bromodomain-containing protein 4 (BRD4) and HPV16 viral load in cervical squamous cell carcinoma and non-cancerous tissues, as well as the effects of BRD4 degradation regent MZ1 on the viral load, proliferation and migration of cervical cancer cells. Real-time fluorescence quantitative PCR showed that the viral load of cervical cancer specimens was significantly higher than that of non-cancer specimens. Immunohistochemical assay showed that BRD4 expression was elevated in cervical cancer specimens (P < 0.001) and in specimens with high viral load (P < 0.0001). Treatment of cervical cancer cells of SiHa, HeLa and CaSki with BRD4 degradation regent MZ1 significantly reduced viral load and inhibited cell proliferation and migration. Nude mouse xenograft tumor confirmed that the tumor volume, and tumor weight of the MZ1-treated mice were significantly lower than those of the control group. Expression of BRD4 and cell proliferation molecule Ki67 in the tumor sections of MZ1-treated mice was significantly decreased, while apoptosis molecule cleaved caspase-3 expression was significantly increased. Moreover, the viral load in the MZ1-treated group was significantly lower than that in the control group. These findings suggest that BRD4 has a potential role in HPV16 viral replication, and MZ1 has a favorable effect in inhibiting viral replication, increasing apoptosis and suppressing the proliferation and migration of cervical cancer cells.

Cinobufagin modulates vasculogenic mimicry and tumor-associated macrophages to inhibit ovarian cancer progression

Ovarian cancer is among the most prevalent malignant tumors affecting women. While conventional therapies like surgery do provide some measure of disease control, they are accompanied by evident side effects that may readily result in drug resistance. Cinobufagin (HCS) is a water-soluble active component extracted from the dried skin of the Bufo gargarizans. Clinical studies have demonstrated its significant anti-tumor effects. Transcriptome sequencing identified Forkhead Box S1 (FOXS1)-related targets, and Western blot analysis evaluated the expression levels of vasculogenic mimicry (VM)-related proteins and pathway proteins after cinobufagin intervention. Immunofluorescence and ELISA were used to detect the effects of cinobufagin on M1 and M2 macrophage markers. Additionally, a co-culture model of Skov3 cells and macrophages was established to study the effects of cinobufagin on tumor-associated macrophage polarization. Cinobufagin significantly inhibited the growth of Skov3 ovarian cancer cells both in vitro and in vivo. Additionally, cinobufagin decreased the expression levels of VM-related proteins, thereby affecting vasculogenesis both in vitro and in vivo. Transcriptomic analysis revealed that the regulation of the FOXS1 gene contributed to this inhibitory effect. In the co-culture system, we found that cinobufagin inhibited IL-4-induced M2 macrophage polarization. Overexpression of FOXS1 in Skov3 cells enhanced the activity of the C-C motif chemokine ligand 2/receptor 2 (CCL2/CCR2) pathway, which was suppressed by cinobufagin, thus affecting the tumor microenvironment. Cinobufagin suppressed vasculogenic mimicry by regulating the FOXS1 gene and inhibited M2 macrophage polarization through the CCL2/CCR2 pathway, thereby affecting the tumor microenvironment.

Differentiation-inducing factor-1 inhibits EMT by proteasomal degradation of TAZ and YAP in cervical and colon cancer cell lines

We previously reported differentiation-inducing factor-1 (DIF-1) activated glycogen synthase kinase-3 (GSK-3) in various mammalian cells. GSK-3 has been proposed to regulate a number of signaling pathway including TAZ/YAP signaling pathway. To clarify the effect of DIF-1 on TAZ/YAP signaling pathway, we examined whether DIF-1 affect the expression levels of TAZ and YAP. We found that DIF-1-induced proteasomal- and GSK-3-dependent degradation of both TAZ and YAP in human cervical cancer cell line HeLa in a time- and dose-dependent manner. As TAZ/YAP signaling pathway is well known to accelerate the epithelial-mesenchymal transition (EMT) of the cancer cell, we examined the effect of TAZ/YAP signaling pathway on EMT-related proteins. Knockdown of TAZ and YAP proteins by siRNA significantly reduced the expression of fibronectin, vimentin, and Snail. We also found that DIF-1 suppressed the expression levels of TAZ/YAP target gene products and EMT-related protein. Further, overexpression of TAZ and YAP attenuated the inhibitory effects of DIF-1 on these protein expressions. Migration and trans-well invasion assays revealed that DIF-1 significantly inhibited HeLa cell migration and invasion. DIF-1-induced proteasomal- and GSK-3-dependent degradation of TAZ and YAP proteins and inhibition of cell migration and invasion were also observed in human colon cancer cell line HCT-116. These results suggest that DIF-1 inhibits the TAZ/YAP signaling pathway via GSK-3 activation. Further, it has been suggested that the inhibition of EMT induced by DIF-1 is involved with the suppression of TAZ/YAP signaling pathway.

Targeting the KRAS-p38-ATF4 axis enhances the therapeutic sensitivity of the peptide PDBAG1 in ovarian cancer via stress response modulation

We previously screened a peptide PDBAG1 that remarkably inhibited triple-negative breast cancer, and found that its target was C1QBP. Recently, C1QBP has been reported as a potential tumor marker in ovarian cancer, which of the mortality rate ranks first among malignant tumors of the female reproductive tract. However, it is unclear whether and how PDBAG1 plays a regulatory role in ovarian cancer. Here, we first found that PDBAG1 definitely inhibited the growth and metastasis of ovarian cancer in vitro and in vivo. PDBAG1 downregulated the protein level of C1QBP and damaged mitochondria in ovarian cancer. Furthermore, we analyzed the overall impact of PDBAG1 on ovarian cancer cells through transcriptomics, and found that KRAS, inflammation and stress-related signals were dramatically activated. The accuracy of the transcriptome sequencing results was also subsequently verified. Moreover, we combined the inhibitors of the classic downstream MAPK signaling pathway of KRAS and the integrated stress inhibitor with PDBAG1, and found that the p38 MAPK inhibitor, Adezmapimod, significantly enhanced the inhibitory effect of PDBAG1 on ovarian cancer and inhibited the upregulation of the crucial stress response transcription factor ATF4 caused by PDBAG1. Collectively, our research results revealed the function and mechanism of the peptide PDBAG1 in ovarian cancer, providing new insights into clinical drug development for ovarian cancer.

Pharmacophore based virtual screening for identification of effective inhibitors to combat HPV 16 E6 driven cervical cancer

Targeting HPV16 E6 has emerged as an effective drug target for the treatment/management of cervical cancer. We utilized pharmacophore-based virtual screening, molecular docking, absorption, distribution, metabolism and excretion (ADME) prediction, and molecular dynamics simulation approach for identifying potential inhibitors of HPV16 E6. Initially, we generated a ligand-based pharmacophore model based on the features of four known HPV16 E6 inhibitors (CA24, CA25, CA26, and CA27) via the PHASE module implanted in the Schrödinger suite. We constructed four-point pharmacophore features viz., three hydrogen bond acceptors (A) and one aromatic ring (R). The common pharmacophore feature further employed as a query for virtual screening against the ASINEX database via Schrödinger suite. The pharmacophore-based virtual screening filtered out top 2000 hits, based on the fitness score. We then applied the high throughput virtual screening (HTVS), standard precision (SP) and extra precision (XP). 1000 compounds were obtained from HTVS docking. Based on the glide score, they were further filtered to 500 hits by employing docking in standard precision mode. Finally, the best four hits and a negative molecule were identified using docking in XP mode. The four lead compounds and a negative molecule were then further subjected to ADME profile prediction by engaging Qikprop module. The ADME properties of the four lead molecules indicate good pharmacokinetic (PK) properties rather than the negative molecule. The binding stability of the HPV16 E6-hit complexes were investigated at a different time scale (100 ns) by using the desmond package and the results were examined using Root Mean Square Deviation (RMSD) and Root Mean Square Fluctuation (RMSF) and it revealed the stability of the protein-ligand complex throughout the simulation. Key residues, CYS 51 and GLN 107, also play a crucial role in enhancing the stability of the protein-ligand complex during the simulation. Furthermore, the binding free energy of the HPV16 E6-leads complexes was analyzed by prime which revealed that the ΔG

Inhibiting the growth of ovarian cancer cells in vitro and in vivo by a small molecular inhibitor targeting La-RNA interactions

To identify small molecules blocking La-RNA interactions by using structural dynamics, molecular biology, and in vivo efficacy experiments. A docking virtual assay on the Chemdiv database was used to screen La binders, and their affinity were measured by surface plasmon resonance (SPR). A novel fluorescence polarization (FP) assay referring to the binding of La protein and 3'UUUOH was established to identify the inhibitors. Their activity on ovarian cancer cell proliferation, apoptosis and cell cycle were evaluated using Cell Counting Kit 8 (CCK8) and flow cytometry assay, respectively. Their in vivo efficacy against ovarian cancer growth were evaluated in a cell line-derived xenograft (CDX) model of A2780 cells. From a total of 20 compounds with high potential binding activity with La protein, two small molecule compounds 4424-1120 and 8017-5932 with relatively stronger inhibition ability on La-RNA interactions were identified. These two compounds shared the same active centers with hydroxyimidazole and hydroxybenzene to interact with La protein through residues ARG57, GLN20 and GLN136. The in vitro assays showed that 4424-1120 and 8017-5932 effectively cause G0/G1 cell cycle arrest, inhibit cell proliferation, reduce cell invasion and promote apoptosis in ovarian cancer cells. In a CDX model on BALB/C Nude mice, we found that the growth rate of the tumor was inhibited by 4424-1120. Our results demonstrated compound 4424-1120 shows good antitumor activity and safety in vitro and in vivo, and it provides a new idea for the discovery of antitumor lead compounds from small drug-like molecules.

Tackling drug resistance in ovarian cancer with epigenetic targeted drugs

Epigenetic dysregulation plays a crucial role in the development and progression of ovarian cancer. Since the first experiment conducted on resistant ovarian cancer cells using demethylating drugs, multiple clinical trials have revealed that epigenetic targeted drugs combined with chemotherapy, molecular-targeted drugs, or even immunotherapy could enhance tumor sensitivity and reverse acquired resistances. Here, we summarized the combination strategies of epigenetic targeted drugs with other treatment strategies of ovarian cancer and discussed the principles of combination therapy. Finally, we enumerated several reasonable clinical trial designs as well as future drug development strategies, which may provide promising ideas for the application of epigenetic drugs to ovarian cancer.

Attenuated Salmonella typhimurium L forms combined with lentivirus shRNA-HOTAIR effectively inhibit tumor growth and metastasis in murine epithelial ovarian cancer

Epithelial ovarian cancer (EOC) is a common gynecological malignant tumor, with a high mortality rate. HOX antisense intergenic RNA (HOTAIR) in lncRNAs is involved in various tumor epithelial-mesenchymal transition (EMT) processes. For seeking better treatment strategies, we studied the effects of attenuated Salmonella typhimurium (ST) L forms combined with lentivirus shRNA-HOTAIR on in vivo tumorigenicity and metastasis of murine EOC cells, and the related anti-tumor mechanisms. Attenuated ST VNP20009 was induced into bacterial L forms by using ceftriaxone. ST L forms were appeared red in Gram staining. Attenuated ST L forms can inhibit the invasion ability of EOC cells in vitro. TUNEL assays showed that attenuated ST L forms combined with lentivirus shHOTAIR can induce more apoptosis of ID8 cells in murine ovarian tumors, compared to the negative control group and only ST or bacterial L forms therapy group. Meanwhile, attenuated ST L forms combined with lentivirus shHOTAIR more effectively inhibited tumor growth and lung metastasis in murine ovarian tumors. The tumorigenicity-related proteins of xenograft tumors detected by immunohistochemistry and qRT-PCR assays showed that attenuated ST L forms combined with lentivirus shHOTAIR can more effectively decrease the protein and mRNA expressions which promote tumor growth and metastasis, such as TGF-β1, ZEB1 and Vimentin. This study confirmed that attenuated ST L forms combined with lentivirus shHOTAIR can more effectively suppress tumor growth and lung metastasis in murine ovarian tumors. Attenuated ST L forms combined with lentivirus shHOTAIR may serve as a better novel biological strategy for bacterial-mediated tumor therapy in EOC.

Tiliroside induces ferroptosis and suppresses tumor growth by synergistically targeting AKR1B1 and modulating iron metabolism in ovarian cancer cells

Ovarian cancer (OC) is a common malignant tumor with the greatest mortality rate among gynecological tumors. Tiliroside (TIL) is a glycosidic dietary flavonoid with various pharmacological activities. The purpose of this study was to investigate the exact mechanism by which TIL eliminates OC cells. In vitro, TIL exerted anti-tumor activities by inducing cell death and inhibiting the invasion and migration of A2780 and OVCAR8 cells. Additionally, the suppressive effect of TIL on OC cells was mainly due to the induction of ferroptosis, as demonstrated by the fact that only ferroprostatin-1 (Fer-1) significantly inhibited the anti-tumor activity of TIL, with the accumulation of ROS, MDA, and Fe Our research revealed that TIL simultaneously targets AKR1B1 and modulates iron metabolism, thereby inducing ferroptosis and improving anti-tumor efficacy. As a novel drug, TIL is promising for OC treatment.

α-Cyperone inhibits the proliferation of human cervical cancer HeLa cells via ROS-mediated PI3K/Akt/mTOR signaling pathway

Cervical cancer is the fourth leading killer of female cancer patients worldwide. Each year more than half a million women are diagnosed with cervical cancer and the disease results in over 300, 000 deaths. α-Cyperone is known as the principal active ingredient in the Cyperus rotundus (Family: Cyperaceae). However, the effects of α-Cyperone on cancers, especially on cervical cancer, are yet to be explored. In the present study, the underlying mechanism of the anti-tumor activity of α-Cyperone against HeLa cells was investigated. The results showed that α-Cyperone inhibited proliferation and induced apoptosis in HeLa cells. Mechanistically, α-Cyperone promoted HeLa cells apoptosis via a mitochondrial apoptotic pathway, which was proved by increased level of intracellular reactive oxygen species (ROS) and upregulated expression of cytochrome c, cleaved caspase-3, PARP, and Bax. Further RNA-sequencing revealed α-Cyperone inhibited the activation of PI3K/Akt/mTOR signaling pathway in HeLa cells, which confirmed by PI3K inhibitor and agonist. The PI3K inhibitor (LY294002) synergized with α-Cyperone in arresting the growth of HeLa cells, whereas the PI3K agonist (IGF-1) abrogated such an effect. Interestingly, the expression of PD-L1 was attenuated by both α-Cyperone and LY294002, while the supplement of IGF-1 rescued the low expression of PD-L1. In conclusion, our results reveal that the inhibitory effect of α-Cyperone on HeLa cell growth is triggered via the ROS-mediated PI3K/Akt/mTOR signaling pathway and closely related to a decline in the PD-L1 expression.

Nanoformulated quinacrine regulates NECTIN-4 domain specific functions in cervical cancer stem cells

NECTIN-4 [a poliovirus receptor-related-4 (PVRL-4) encoded gene] has vital roles in cancer proliferation, metastasis and angiogenesis. It possesses three different domains and it is predicted that they have different roles in cancer but the structure-function relationship is still unknown and hence carrying out a detailed study to elucidate the domain-specific functions of NECTIN-4 in cancer is necessary. Using 5-Fluouracil-resistant cervical cancer stem cells (PEMT-5FU-R-MC) and different NECTIN-4 domain-specific constructs, different domains of NECTIN-4 were over-expressed in PEMT-5FU-R-MC cells. Biochemical assays like comet, γ-H2AX immunofluorescence, western blot, in vitro tube formation, gelatin zymography, in ovo CAM assay, etc. were used to delineate the function of each domain of NECTIN-4 in cancer and their regulation by nano-formulated quinacrine (NQC). Endo-domain (lacking extracellular region corresponding to aa 30-347) and ecto-domain (lacking signal peptide and cytoplasmic region corresponding to aa 1-29 and 348-509, respectively) of NECTIN-4 were largely overexpressed in nucleus and cytoplasm, respectively. Endo-domain translocates into nucleus by physically interacting with IMPORTIN-α2, activates the DNA repair and enhances cell growth, whereas ecto-domain specifically activates angiogenesis by modulating representative angiogenic markers, inducing in vitro tube formation and in ovo blood vessel formation. Full-length NECTIN-4 (aa 1-509) was overexpressed in both nucleus and cytoplasm and modulated both DNA repair and angiogenesis. NQC down-regulated these phenomena by modulating the endo-domain and ecto-domain of NECTIN-4. Thus, current study suggested that endo-domain of NECTIN-4 translocated into nucleus and increased the DNA repair and ecto-domain of NECTIN-4 enhanced the angiogenesis, whereas NQC inhibits these processes.

METTL3 regulates the malignancy of cervical cancer via post-transcriptional regulation of RAB2B

Cervical cancer is one of the leading causes of cancer death in women worldwide. While molecular mechanisms of initiation and cervical carcinogenesis are not well studied. Our data showed that the expression of Methyltransferase-like 3 (METTL3) was upregulated in cervical tumor tissues as compared with normal tissues. Its expression was associated with poor prognosis of cervical cancer. Knockdown of METTL3 can suppress the proliferation of cervical cancer cells. The expression of METTL3 was significantly correlated with the expression of RAB2B, one member of RAS oncogene family. Over expression of RAB2B can significantly attenuate sh-METTL3-suppressed cell proliferation. Mechanistically, METTL3 can increase the mRNA stability of RAB2B via an IGF2BP3-dependent manner. Collectively, METTL3 can trigger growth of cervical cancer cells via upregulation of RAB2B. It indicated that METTL3 might be a potential target for cervical cancer therapy.

Purified compounds from marine organism sea pen induce apoptosis in human breast cancer cell MDA-MB-231 and cervical cancer cell Hela

Marine organisms are an important source of chemical compounds which are appropriate for use as therapeutic agents. Among them, Sea pens produce valuable chemical compounds being used as anti-cancer drugs. The aim of this study was to investigate anti-cancer property of extracted and purified compounds from marine organism Sea pen and evaluate their effects on inducing of apoptosis. The extracts were prepared from dried colony of Virgularia gustaviana. The compounds (3β)-Cholest,5en,3ol (cholesterol) (15 mg), Hexadecanoic acid (2.5 mg) and 2-Hexadecanol (10.7 mg) were identified by GC-MS and NMR. The cytotoxic effects of the compounds were evaluated on Hela and MDA-Mb-231 human cancer cell lines with MTT assay. Immunocytochemistry and Western Blot analyses were used to evaluate the expression of apoptosis related markers Caspase 3, Caspase 8, Bax and BCL2 in cancer cells after treating with three compounds. The purified compounds reduced viability of human breast cancer cell line MDA-MB-231 and human cervical cancer cell line Hela concentration-dependently. 2-Hexadecanol reduced significantly the viability of both cancer cell lines in comparison to the other purified compounds. Treatment of cancer cells with the three purified compounds increased the expression of caspase-3, caspase-8 and Bax proteins and decreased the relative Bcl-2/Bax ratio, demonstrating induction of apoptosis as possible mechanism of action. According to the results, three purified compounds inhibit the growth of cancer cells by inducing of apoptosis pathway; an effect which needs to be further investigated in the future studies.

Vorinostat upregulates MICA via the PI3K/Akt pathway to enhance the ability of natural killer cells to kill tumor cells

Vorinostat has good therapeutic efficacy against primary cutaneous T-cell lymphoma in the refractory stage. However, the molecular mechanism by which it inhibits solid tumors has not been clarified. To investigate the tumor inhibitory mechanism of vorinostat in cervical cancer, this study used Cell Counting Kit-8, flow cytometry, cell invasion and migration assays and the wound healing assay to evaluate the effects of vorinostat on cervical cancer cell proliferation, apoptosis, cell cycle, migration, and invasion. Real-time quantitative PCR and immunoblotting were used to detect gene and protein expression, respectively, of major histocompatibility class I-related chain A, phosphoinositide 3-kinase, phosphorylated PI3K p55 (Tyr199), and p-Akt (Ser473). The lactate dehydrogenase cytotoxicity assay was used to evaluate the ability of natural killer-92 cells to lyse cervical cancer cells. A xenograft nude mouse model was established to analyze the anti-tumor effect of vorinostat in vivo. Our results showed that vorinostat inhibited the proliferation, migration, and invasion of cervical cancer cells. Vorinostat also induced apoptosis and cell-cycle arrest in the S phase, inhibited PI3K (p110α), p-PI3K p55 (Tyr199), and p-Akt (Ser473) protein expression and upregulated MICA expression in vitro and in vivo, and promoted NK-92 cell-mediated cervical cancer cell lysis. The ability of vorinostat to upregulate MICA expression in cervical cancer cells was related to PI3K/Akt signaling. In brief, vorinostat upregulated MICA through the PI3K/Akt pathway and enhanced the sensitivity of cervical cancer cells to the NK cell-mediated cytolytic reaction. The results of this study demonstrate that vorinostat has anti-solid tumor effects on cervical cancer.

Apoptotic, necrotic, and antiproliferative activity of diosgenin and diosgenin glycosides on cervical cancer cells

(25R)-spirost-5-en-3β-ol, also known as diosgenin (DSG), exerts antiproliferative activity on diverse cell lines, induces apoptosis, and acts as a chemopreventative agent. However, the relationship between DSG glycosides and apoptotic, necrotic, and antiproliferative activity remains unclear. It is in this regard that we report the antiproliferative, necrotic, and apoptotic activities of DSG and its glycoside derivatives: (25R)-spirost-5-en-3β-yl O-β-D-glucopyranoside (3GD), (25R)-spirost-5-en-3β-yl O-α-L-rhamnopyranosyl-(1 → 4)-β-D-glucopyranoside (3GRD); and (25R)-spirost-5-en-3β-yl O-α-L-rhamnopyranosyl-(1 → 2)-O-[α-L-rhamnopyranosyl-(1 → 4)]-β-D-glucopyranoside), also known as dioscin (DSC), in in vitro assays of cervical HeLa and CaSki cancer cells. The results demonstrated that DSG glycosidic derivatives preserved their antiproliferative activity. However, in both cancer cell lines, 3GD and 3GRD were less potent than DSG, while DSC was more potent than DSG. With respect to necrotic activity, all tested compounds showed no or low activity on the two cervical cancer cell lines. Regarding apoptosis, the results showed that DSG glycosides were better apoptosis-inducers than DSG, suggesting that glucose and rhamnose residues play a central role in enhancing the apoptotic activity of DSG. Finally, DSG and its glycosidic derivatives were shown to affect the proliferative potential of lymphocytes (non-tumour cells) to a lesser extent than cancer cells, suggesting that these compounds have selective action. In conclusion, the results indicate that DSG and its glycosidic derivatives are promising anticancer compounds since they are compounds with low necrotic activity and selective action.

Hederagenin suppresses ovarian cancer via targeting mitochondrial fission through dynamin-related protein 1

A triterpenoid isolated from the plant Hedera helix, hederagenin was discovered to have anti-cancer, anti-inflammatory, anti-depressant and anti-fibrosis properties both in vivo and in vitro. In this study, the relationship between mitochondrial fission and hederagenin-induced apoptosis in ovarian cancer (OC) was investigated and the underlying mechanisms were deciphered. Hederagenin's cytotoxicity on OC cells was analyzed using colony formation and CCK-8 assays. The effect of hederagenin on OC cells was also verified by a mouse xenograft tumor model. Flow cytometric analysis was conducted to examine hederagenin's effects on mitochondrial membrane potential, apoptosis, and cell cycle OC cells. MitoTracker Red (CMXRos) staining was performed to observe the mitochondrial morphology. The protein levels of Bak, Bcl-2, Caspase 3, Caspase 9, Cyclin D1 and Bax were measured by Western blot. This study found that hederagenin could suppress the in vivo and in vitro SKOV3 and A2780 cell proliferation in an effective manner. Besides, hederagenin altered the mitochondrial membrane potential, induced S-phase and G0/G1-phase arrest, mitochondrial morphology changes, and apoptosis in OC cells. Additionally, our findings further demonstrated that hederagenin changed the mitochondrial morphology by suppressing dynamin-related protein 1 (Drp1), a crucial mitochondrial division factor. Moreover, Drp1 overexpression could reverse hederagenin-induced apoptosis, whereas the Drp1 knockdown had the opposite effect. Furthermore, hederagenin may trigger BAX mitochondrial translocation and apoptosis in OC cells. These results provided a novel perspective on the relationship between the modulation of mitochondrial morphology and the suppression of ovarian cancer by hederagenin.