Tissue and Cell

POU6F1 inhibits cervical cancer progression by activating MAOB to block the ERK1/2 pathway

Cervical cancer (CC) ranks as the fourth common malignancy among women. This study investigated the mechanism by which POU6F1 mediates CC progression through MAOB. MAOB expression was detected in CC tissues and cell lines. The effects of MAOB overexpression on cell proliferation, migration, and invasion were observed using CCK-8, EdU, colony formation, wound healing, and Transwell assays. Western blot was conducted to assess the extent of ERK1/2 phosphorylation. CC cell lines were treated with an ERK1/2 phosphorylation activator based on OE-MAOB to verify the effects of the ERK1/2 phosphorylation activator on CC cell growth. Transcription factors binding to the MAOB promoter were predicted. POU6F1 expression in cells was detected. Gene interference of OE-POU6F1 was performed, and the transcriptional relationship between POU6F1 and MAOB was confirmed through dual-luciferase and ChIP experiments. Based on OE-POU6F1, MAOB was knocked down to verify that POU6F1 affected CC development via MAOB in vitro and in vivo. MAOB was downregulated in CC tissues (versus adjacent tissues) and cell lines (versus human cervical epithelial cells). MAOB overexpression repressed the proliferation, migration, and invasion of CC cells and suppressed the phosphorylation of ERK1/2. Treatment with an ERK1/2 phosphorylation activator weakened the repressive effect of MAOB overexpression on CC cell growth. POU6F1 was poorly expressed in CC cells, and POU6F1 transcriptionally activated MAOB. Combined knockdown of MAOB reversed the inhibited cell behaviors and tumor growth observed when POU6F1 was overexpressed alone. POU6F1 inhibits CC progression by activating MAOB transcription and suppressing the ERK1/2 signaling activation.

Targeting Siglec-15 mediates mitochondrial retrograde regulation of cervical cancer development

Cervical cancer (CCA) is the predominant cause of fatalities from gynecologic malignancies, with metastasis responsible for 80 % of cancer-related mortalities. This study preliminarily examined the involvement of Sialic Acid Binding Ig Like Lectin 15 (Siglec-15) in the development of CCA and its probable mechanisms. We assessed the capacity of Siglec-15 to modulate CCA progression by establishing knockdown and overexpression Siglec-15 cell lines, supplemented with animal models, using both in vivo and in vitro dual investigations. Our findings indicate that Siglec-15 is significantly expressed in CCA cell lines and is intimately associated with the proliferation, migration, and invasion capabilities of CCA cells, as well as mitochondrial ROS homeostasis. The suppression of Siglec-15 expression markedly reduced tumor growth in mice, potentially due to Siglec-15's role in regulating the Mitogen-Activated Protein Kinase (MAPK) signaling pathway, which mediates the retrograde regulation of mitochondrial ROS homeostasis. Siglec-15 may emerge as a novel therapeutic target and prognostic marker for patients with CCA.

Anticancer activity of Momordica charantia-derived extracellular vesicle-like particle against cervical cancer

To investigate the function and possible mechanisms of Momordica charantia-derived extracellular vesicle-like particle (MC-EVLP) in cervical cancer in vitro and in vivo. The half-maximal inhibitory concentration (IC50) of MC-EVLP in HeLa and C33A cells were calculated. In this study, colony formation, BrdU staining, cell migration, cell apoptosis, TUNEL staining, malondialdehyde, glutathione and ferrous iron levels, cell viability, and PCNA, CyclinD1, N-cadherin, Vimentin, E-cadherin, matrix metalloproteinase 9, Bcl-2, Bax, p-Akt, GPX4, ACSL4, and transferrin receptor levels were evaluated. In a HeLa cell line xenograft tumor model, the effects of MC-EVLP on the tumor growth rate and volume and the levels of Ki67 and N-cadherin were also evaluated. The IC50 values of MC-EVLP were determined to be 59 µg/ml in HeLa cells and 68 µg/ml in C33A cells. MC-EVLP significantly reduced colony formation, the percentage of BrdU-positive cells, and the levels of the proliferative markers PCNA and Cyclin D1 in both cell lines. Additionally, MC-EVLP inhibited cell migration and decreased N-cadherin, vimentin, and matrix metalloproteinase 9 levels. Apoptosis was increased, as evidenced by an increased apoptosis rate, an increased ratio of TUNEL-positive cells, a reduced Bcl-2/Bax ratio, and decreased p-Akt levels. Moreover, MC-EVLP increased lipid peroxidation, downregulated glutathione peroxidase 4 expression, and significantly increased the expression of ACSL4 and transferrin receptor, with Fer-1 partially reversing these effects. In vivo, the intraperitoneal injection of MC-EVLP significantly suppressed tumor growth and decreased the expression of Ki-67 and N-cadherin. MC-EVLP demonstrated considerable cytotoxic effects on cervical cancer. They promote apoptosis by activating the Bcl-2/Bax/p-Akt pathway and induce ferroptosis by upregulating GPX4 and iron-mediated mechanisms. These findings suggest that MC-EVLP have potential as anticancer agents.

OTX1 silencing suppresses ovarian cancer progression through inhibiting the JAK/STAT signaling

The aim of our study was to investigate the roles and the underlying mechanisms of orthodenticle homolog 1 (OTX1) in ovarian cancer. OTX1 expression was obtained from TCGA database. OTX1 expression in ovarian cancer cells was detected using qRT-PCR and western blot assay. The cell viability and proliferation was detected by CCK-8 and EdU assays. Cell invasion and migration were detected by transwell assay. Flow cytometry was utilized to determine cell apoptosis and cycle. In addition, western blot assay was used to detect the expression of cell cycle related protein (Cyclin D1 and p21), epithelial-mesenchymal transition (EMT) related protein (E-cadherin, N-cadherin, Vimentin, and Snail), apoptosis related protein (Bcl-2, Bax, and cleaved caspase-3), and JAK/STAT pathway related protein (p-JAK2, JAK2, STAT3, and p-STAT3). OTX1 was highly expressed in ovarian cancer tissues and cells. OTX1 silencing blocked the cell cycle and repressed cell viability, proliferation, invasion, and migration, while OTX1 silencing facilitated the apoptosis of OVCAR3 and Caov3 cells. OTX1 silencing increased the protein levels of p21, E-cadherin, Bax, and cleaved caspase-3, while the protein levels of Cyclin D1, Bcl-2, N-cadherin, Vimentin, and Snail were decreased by OTX1 silencing. Furthermore, OTX1 silencing suppressed the protein levels of p-JAK2/JAK2 and p-STAT3/STAT3 in OVCAR3 and Caov3 cells. Moreover, overexpression of OTX1 promoted cell proliferation and invasion and inhibited apoptosis in Caov3 cells, but AG490 (an inhibitor of JAK/STAT pathway) reversed the influences on cell biological behavior induced by overexpression of OTX1. OTX1 silencing repressed ovarian cancer cell proliferation, invasion, and migration and induced cell apoptosis, which might be involved in JAK/STAT signaling pathway. OTX1 may be considered as a novel therapeutic target for ovarian cancer.

Knockdown of CENPU inhibits cervical cancer cell migration and stemness through the FOXM1/Wnt/β-catenin pathway

Currently, the clinical outcome of cervical cancer (CC) is still undesirable, and it is urgent to explore more treatment strategies for CC. In this study, the effects of CENPU on migration and stemness of CC was studied. The CENPU expression were retrieved from The Cancer Genome Atlas (TCGA). The effects of CENPU on the viability and proliferation of cells were evaluated by CCK-8 assay and colony formation assay. Wound healing assay and invasion assay were chosen to assess migration and invasion of cells. Tumorsphere-forming assay was applied for testing the stemness. Western blot analysis was applied for assessing the level of CENPU, Nanog, Oct4, FOXM1, β-catenin, c-myc and MMP-7. The tumor sizes and volumes were also measured. The TCGA data and WB assay suggested that CENPU was upregulated in CC. CENPU knockdown would inhibit the viability of CC cells and prohibit the migration and invasion of cells. Tumorsphere-forming assay and WB results suggested that CENPU silencing decreased the sphere formation rate and the expression of Nanog and Oct4. Moreover, CENPU knockdown suppressed the expression of FOXM1, β-catenin, c-myc and MMP-7 by WB. In vivo study demonstrated that CENPU knockdown inhibited the growth of CC, indicated by reduced sizes and volumes of CC. In summary, our results suggested that knockdown of CENPU inhibited CC migration and stemness through the FOXM1/Wnt/β-catenin pathway.

Ultrasound microbubble-mediated delivery of ANLN silencing-repressed EZH2 expression alleviates cervical cancer progression

Ultrasound-targeted microbubble destruction (UTMD) is a new gene therapy method that uses ultrasound and microbubbles carrying target genes to achieve gene transfection. However, whether UTMD-mediated ANLN silencing transfection helps to restrain the growth of cervical cancer (CC) is obscure. ANLN level in tumor tissues, adjacent tissues, and cells was tested using the database, qRT-PCR, and western blot. The optimal concentration of SF6 was determined by MTT assay. Mechanical index (MI) was selected by flow cytometry. After transfection with liposome or UTMD-mediated liposome, cell function experiments, qRT-PCR, and western blot were employed to assess CC cell biological behaviors and EZH2 level. Epithelial-mesenchymal transition (EMT)-related marker and apoptosis-related marker expressions were examined utilizing qRT-PCR and western blot. 10% SF6 and MI of 0.28 were selected for subsequent tests. ANLN was highly expressed in CC and cells. The transfection efficiency of the UTMD-siANLN group was higher than that of the L-siANLN group. Moreover, the repression of UTMD-siANLN on CC cell malignant phenotypes was stronger than L-siANLN. UTMD-siANLN attenuated EZH2 expression in CC cells. The modulatory role of UTMD-siANLN on EMT- and apoptosis-related markers was reversed by EZH2 overexpression. UTMD can improve the efficiency of siANLN transfection into CC cells to induce suppression of CC cell malignant phenotypes, which may become a new target of gene therapy for CC.

RACGAP1 promotes proliferation and cell cycle progression by regulating CDC25C in cervical cancer cells

RACGAP1 (Rac GTPase-activating protein 1) is correlated with tumor aggressiveness and poor prognosis, but the role of RACGAP1 in cervical cancer has not been fully reported. Analysis of RACGAP1 expression data in cervical cancer from the Cancer Genome Atlas (TCGA) database was carried out by GEPIA and UALCAN websites. In addition, the UALCAN database was used to identify the RACGAP1 positively correlated genes, which were used for the enrichment analysis. qRT-PCR, immunohistochemistry, western blot, and immunofluorescence were utilized to measure RACGAP1 expression in tissues and cells. Western blot, flow cytometry, MTT, and colony formation assays were applied to assess the effects of RACGAP1 on cell cycle, growth and viability in cervical cancer. Through bioinformatics analysis, we found that the level of RACGAP1 was aberrantly increased in cervical cancer, which was confirmed in cervical cancer tissues and cells. RACGAP1 associated genes, including CDC25C, were mainly enriched in cell cycle pathway, and RACGAP1 expression was negatively associated with CDC25C expression. RACGAP1 overexpression was related to patient's poor prognosis and promoted cervical cancer cell proliferation. Furthermore, RACGAP1 knockdown decreased the level of CDC2, p-CDC2, CDC25C, and Cyclin B1, inhibited proliferation and delayed cell cycle progression in cervical cancer cells. In mechanism, overexpression of CDC25C attenuated RACGAP1 knockdown-mediated cell growth inhibition and cell cycle arrest. Taken together, this study demonstrated that RACGAP1 was overexpressed in cervical cancer, and downregulation of RACGAP1 could inhibit the cervical cancer cell proliferation and cell cycle progression through regulating CDC25C expression.

Smad7 drives cisplatin resistance in ovarian cancer via a PRMT5-dependent mechanism

Acquired cisplatin resistance poses a major challenge in ovarian cancer management. This study investigated the role of protein arginine methyltransferase 5 (PRMT5) in this context. Using cisplatin-resistant ovarian cancer cell lines (A2780/DDP and SKOV3/DDP), we found that PRMT5 knockdown significantly inhibited cell proliferation, colony formation, migration, and invasion, while promoting apoptosis. Mechanistically, co-immunoprecipitation assays revealed that PRMT5, in complex with MEP50, interacts with and specifically methylates Smad7 at the R57 site in vitro. This methylation event was essential for activating the STAT3 signaling pathway and driving the observed malignant phenotypes. Consistent with a key oncogenic role for Smad7, in vivo knockdown of Smad7 in a xenograft mouse model markedly suppressed tumor growth and downregulated markers of proliferation (Ki67), invasion (MMP9, N-cadherin), while upregulating the tumor suppressor E-cadherin. In conclusion, our work identifies Smad7 as a critical driver of cisplatin resistance in vivo and delineates a novel in vitro mechanism whereby PRMT5 promotes oncogenic signaling through R57 methylation of Smad7. This PRMT5-Smad7 axis presents a promising therapeutic target for overcoming cisplatin resistance in ovarian cancer.

Scopoletin inhibits ovarian cancer progression by reducing glycolysis via the EGFR-AKT pathway

This study aimed to investigate the antitumor effects and mechanisms of scopoletin in chemoresistant ovarian cancer (OC) cells. Cell viability, migration, invasion, and the cell cycle were assessed. The effect of scopoletin on aerobic glycolysis was determined by measuring glucose uptake, lactate, ATP, extracellular acidification rate (ECAR), and oxygen consumption rate (OCR). Bioinformatics was used to analyze the relationship between scopoletin and epidermal growth factor receptor (EGFR). The levels of p-EGFR and p-AKT were measured using western blotting. In vivo xenograft models were used to validate the role of scopoletin. These results revealed that scopoletin inhibited the viability, migration, invasion, and colony formation of chemoresistant OC cells. It induced concentration-dependent G1/S phase arrest and reversed aerobic glycolysis by reducing glucose uptake, lactate production, ATP levels, and ECAR while increasing the OCR. Scopoletin targets the EGFR-AKT axis and downregulates p-EGFR and p-AKT expression. Crucially, the combination of scopoletin and the EGFR inhibitor lapatinib synergistically inhibited cellular metabolic reprogramming and proliferation and significantly enhanced tumor growth inhibition in mice compared to single-drug treatment. In conclusion, scopoletin suppresses chemoresistant OC progression and metastasis by targeting the EGFR-AKT pathway. The synergistic effect of scopoletin and lapatinib demonstrates a promising therapeutic strategy for overcoming OC chemoresistance.

Cell-type dependent effect of 3D collagen matrix on cancer cell resistance to suboptimal conditions: the case of serum deprivation, glucose starvation, and hypoxia

The extracellular matrix (ECM) and its primary chemical components, including collagen, play a pivotal role in carcinogenesis and tumor progression. The ECM actively regulates cell proliferation, migration, and, importantly, resistance to various adverse factors. It is widely recognized as a key factor in modifying the resistance of tumor cells to various treatment modalities and cytotoxic compounds. However, the role of the ECM in tumor cell adaptation to nutritional deficiencies and hypoxic conditions remains significantly less studied. Since it is generally accepted that tumor cells resistance increases when cultured in a three-dimensional matrix, we sought to experimentally test the universality of this statement. In this work, we analyzed the responses of tumor cells with varying origins and proliferative activities, including human bladder carcinoma, epidermoid carcinoma, and ovarian carcinoma, to deprivation of serum, glucose and oxygen. We compared cell resistance to suboptimal conditions when cultured in a monolayer on tissue culture (TC)-treated polystyrene, on collagen-coated surfaces, or within a three-dimensional hydrogel composed of collagen type I. All three cell lines were stably transfected with fluorescent protein genes. To register the cell growth dynamics, we used a fluorescence-based technique that allows long-term quantitative observations without disrupting the hydrogel. The analyzed cell lines demonstrated different patterns of relative sensitivity to suboptimal conditions. We revealed that the direction and intensity of the collagen matrix effect depend on the cell type. Slowly proliferating ovarian carcinoma cells showed no noticeable changes in their behavior when cultured in a gel compared to a monolayer. In the case of bladder carcinoma, we registered predominantly resistance-stimulating effect of the collagen matrix, but it was significant only under serum deprivation. The most pronounced effect of collagen was registered for epidermoid carcinoma. Importantly, this effect was ambivalent: gel-embedded cells demonstrated significantly enhanced resistance to serum deprivation, but, at the same time, they were more responsive to glucose starvation and hypoxic conditions. We attribute the registered phenomenon to the individual characteristics of tumor cells with different origins and metabolic activities.

Anti-neoplastic effect of heterophyllin B on ovarian cancer via the regulation of NRF2/HO-1 in vitro and in vivo

Heterophyllin B (HB) is a cyclic peptide with anti-neoplastic effect on many cancers. However, its effect and mechanism of action in ovarian cancer cells are still unknown. The primary objective of this study was to assess the impact of HB on the proliferation of ovarian cancer (OC) cells and delve into the underlying mechanisms involved. We performed CCK-8 assays, HE staining, KI67 staining, clonogenic formation assays, Annexin V-FITC/PI staining, tumor invasion assays, and migration assays to detect the effects of HB on cell viability, proliferation, apoptosis, migration, and invasion in ovarian cancer cells. Additionally, real-time fluorescent quantitative PCR (qPCR) and Western blotting were utilized for verification. The expression of NF-E2-related factor 2 (NRF2) and heme oxygenase 1 (HMOX1/HO-1) signaling molecules was detected using qPCR and Western blotting. A specific inducer, Hemin, was used to activate HO-1 and Nrf2 overexpression, in order to verify the pharmacological mechanism of HB on ovarian cancer cells. The binding relationship between HB and NRF2 was investigated through molecular docking. HB treatment inhibited the viability of OC cells, meanwhile it showed suppressive effect on the proliferation, migration, and invasion of OC cells, Meanwhile, HB could promote the apoptosis of tumor cells. For the mechanisms, we found that HB treatment could significantly down-regulate the levels of NRF2/HO-1. Consistent with the results of in vitro experiments, administration of HB significantly delayed tumor growth in OVCAR8 xenografted nude mice, and inhibited the expression of Ki67, Nrf2 and HO-1. This study demonstrated that HB had anti-neoplastic effect on OC by inhibiting Nrf2/HO-1 signaling pathway and may be a potential drug for the treatment of OC.

CISD2 downregulation participates in the ferroptosis process of human ovarian SKOV-3 cells through modulating the wild type p53-mediated GLS2/SAT1/SLC7A11 and Gpx4/TRF signaling pathway

CISD2 and ferroptosis participate in cancer development, but are rarely reported in ovarian cancer. This study aimed to clarify interaction between CISD2 and ferroptosis and evaluate related mechanisms. si-CISD2, wt-p53 and mut-p53 lentiviruses were transfected into SKOV-3 cells. CISD2 and p53 (wild/mutant p53) gene transcriptions were evaluated by RT-PCR. Cell viability, invasion ability, and migration capacity were determined. Expressions of ferroptosis-associated CISD2, p53, elastin, β-catenin and levels of Gpx4 and TRF were examined. CISD2 downregulation (si-CISD2) has a significant inhibitory effect on cell activity and exerts a synergistic effect with p53. si-CISD2 and Wt-p53 markedly inhibited SKOV-3 invasion and migration capacity, compared to the downregulation control (si-NC) and overexpression control (ov-NC) group (p < 0.001). p53 expression was increased significantly in si-CISD2 treated SKOV-3, compared to si-NC treated cells (p < 0.05). si-CISD2 markedly decreased elastin and β-catenin expression compared to the si-NC and ov-NC group (p < 0.001). si-CISD2 modulated ferroptosis-associated molecules (CDKN1A, GLS2, SAT1, SLC7A11), decreased Gpx4 and increased TRF levels in SKOV-3. si-CISD2 and Wt-p53 played an obvious synergistic role in regulating ferroptosis-associated molecules and Gpx4/TRF pathway molecules. In conclusion, CISD2 downregulation was involved in ferroptosis process of SKOV-3 cells. This effect of CISD2 was mediated by wild-type p53-associated GLS2/SAT1/SLC7A11 and Gpx4/TRF pathway.

Lysine demethylase 3A in hypoxic macrophages promotes ovarian cancer development through regulation of the vascular endothelial growth factor A/Akt signaling

Hypoxia is a vital feature of the tumor microenvironment of OC. Previous evidence exposes that tumor-associated macrophages (TAMs) are connected with the development of ovarian cancer (OC), whereas the accurate regulatory mechanism of hypoxic macrophages regulating tumor advancement remains unclear. Herein, we examined whether the lysine demethylase 3 A (KDM3A) in hypoxic macrophages expedited the development of OC cells. The contents of hypoxia inducible factor-1α (HIF-1α), CD163, CD80, KDM3A, and p-Akt/Akt were detected by western blot. Genomic Spatial Event 4630, Molecular Signatures Database, and Comparative Toxicogenomics Database were utilized for correlated gene prediction. The OC cells viability was scrutinized by cell counting kit-8 assay. The cell proliferation was inspected by 5-Ethynyl-2'-deoxyuridine assay. The vascular endothelial growth factor A (VEGF) level was detected by Enzyme-linked immunosorbent assay. M2 polarization of TAMs was associated with poor prognosis in sufferers with OC. The OC sufferers with high level of CD163 or low level of CD80 were linked with poor overall survival and disease specific survival. Hypoxia induced THP-1-derived macrophages M2 polarization. KDM3A was high-expressed in hypoxia induced macrophages. Upregulated KDM3A in hypoxic macrophages facilitated OC cell proliferation. KDM3A upregulation in hypoxic macrophages stimulated Akt signaling activation in OC cells. KDM3A in hypoxic macrophages promoted VEGF secretion to activate Akt signaling in OC cells. VEGF inhibition or Akt signaling inactivation reversed the effects of KDM3A in hypoxic macrophages on OC cells viability and proliferation. The KDM3A content and M2 polarization were enhanced in hypoxic macrophages, and KDM3A in hypoxic macrophages promoted OC development through regulation of the VEGF/Akt signaling pathway.

Collagen I dysregulation is pivotal for ovarian cancer progression

As a principal matrisomal protein, collagen is involved in the regulation of the structural framework of extracellular matrix (ECM) and therefore is potentially crucial in determining the biophysical character of the ECM. It has been suggested that collagen architecture plays a role in ovarian cancer development, progression and therapeutic responses which led us to examine the collagen morphology in normal and cancerous ovarian tissue. Also, the behaviour of ovarian cancer cells cultured in four qualitatively different collagen gels was investigated. The results here provide evidence that collagen I morphology in the cancerous ovary is distinct from that in the normal ovary. Tumour-associated collagen I showed streams or channels of thick elongated collagen I fibrils. Moreover, fibril alignment was significantly more prevalent in endometrioid and clear cell cancers than other ovarian cancer subtypes. In this work, for the first-time collagen I architecture profiling (CAP) was introduced using histochemical staining, which distinguished between the collagen I morphologies of ovarian cancer subtypes. Immunohistochemical examination of ovarian normal and cancerous tissues also supported the notion that focal adhesion and Rho signalling are upregulated in ovarian cancers, especially in the high-grade serous tumours, as indicated by higher expression of p-FAK and p190RhoGEF. The results also support the concept that collagen I architecture, which might be collagen I concentration-dependent, influences proliferation in ovarian cancer cells. The study provides evidence that modification of collagen I architecture integrity is associated with ovarian cancer development and therapeutic responses.

Cervical cell multi-classification algorithm using global context information and attention mechanism

Cervical cancer is the second biggest killer of female cancer, second only to breast cancer. The cure rate of precancerous lesions found early is relatively high. Therefore, cervical cell classification has very important clinical value in the early screening of cervical cancer. This paper proposes a convolutional neural network (L-PCNN) that integrates global context information and attention mechanism to classify cervical cells. The cell image is sent to the improved ResNet-50 backbone network to extract deep learning features. In order to better extract deep features, each convolution block introduces a convolution block attention mechanism to guide the network to focus on the cell area. Then, the end of the backbone network adds a pyramid pooling layer and a long short-term memory module (LSTM) to aggregate image features in different regions. The low-level features and high-level features are integrated, so that the whole network can learn more regional detail features, and solve the problem of network gradient disappearance. The experiment is conducted on the SIPaKMeD public data set. The experimental results show that the accuracy of the proposed l-PCNN in cervical cell accuracy is 98.89 %, the sensitivity is 99.9 %, the specificity is 99.8 % and the F-measure is 99.89 %, which is better than most cervical cell classification models, which proves the effectiveness of the model.

Liposome-engineered therapeutics: A promising frontier in ovarian cancer treatment

Ovarian cancer remains a leading cause of gynecological cancer-related mortality, with epithelial ovarian carcinoma, germ cell tumors, and stromal tumors being the most prevalent types. The disease is classified based on the Fédération Internationale de Gynécologie et d'Obstétrique (FIGO) staging system, and its incidence increases with age. Standard treatment strategies, including taxanes, platinum-based drugs, monoclonal antibodies, and poly (ADP-ribose) polymerase (PARP) inhibitors, as well as emerging gene and immunotherapies, often suffer from significant adverse effects and limited efficacy. To overcome these challenges, advanced drug delivery systems are essential for enhancing therapeutic outcomes while minimizing toxicity. Liposomes have emerged as a promising nanocarrier for targeted drug delivery in ovarian cancer therapy, offering improved drug stability, prolonged circulation time, and enhanced tumor-specific accumulation. Functional modifications, such as ligand conjugation and combination therapies, further optimize their therapeutic potential. This review discusses recent advancements in liposome-based drug delivery for ovarian cancer, highlighting their benefits, challenges, and future directions in improving treatment efficacy.

Ovulation-derived extracellular vesicles exhibit sustained oncogenic influence on the exposed fallopian tube fimbrial cells after drainage into peritoneal cavity

Ovulation is known to damage fallopian tube epithelial (FTE) cells, promote cellular transformation, and contribute to the development of high-grade serous ovarian carcinoma (HGSC). While ovulatory follicular fluid-derived extracellular vesicles (EVs) have been shown to possess cell-transforming activity, it remains unclear whether these EVs persist in the peritoneal cavity after ovulation, potentially prolonging their exposure and enhancing their transformative effects on fallopian tube epithelial (FTE) cells. In this study, we collected follicular fluid (FF) and peritoneal fluid (PF) from women before and after ovulation, and investigated the oncogenic potential of ovulation-derived EVs on FTE cells using an anchorage-independent colony growth assay. We found that post-ovulatory PF exhibits significantly higher cell transformation activity compared to pre-ovulatory PF. This heightened activity correlates with an increased concentration of EVs and protein content in post-ovulatory PF. FF samples obtained from different ovulatory follicles of the same patient demonstrated consistent transformation activity, and FF- or post-ovulatory PF derived EVs retained this transforming capacity across FTE cells at varying stages of transformation. Our study reveals a novel mechanism by which ovulation may contribute to FTE transformation through the persistent oncogenic effects of EVs released into the peritoneal microenvironment. This finding provides new perspectives and directions for future cancer prevention, treatment, and potential diagnostic biomarker research.

CDCA3 exhibits a role in promoting the progression of ovarian cancer

Ovarian cancer (OC) is one of the common gynecological malignant tumors. Cell division cycle-associated protein-3 (CDCA3) is involved in the regulation of cell cycle progression. The role of CDCA3 in OC was explored in this study. The expression of CDCA3 in OC was evaluated in the Gene Expression Omnibus (GEO) database and further verified by qRT-PCR and Western blot (WB). Subsequently, we established lentivirus-mediated CDCA3 knockdown in OC cell lines HO-8910 and A2780. The biological roles of CDCA3 on proliferation, sensitivity to cisplatin, apoptosis, migration and tumor formation of OC were investigated using loss-of-function assays. CDCA3 was frequently up-regulated in OC. Knockdown of CDAC3 inhibited proliferation and migration, and enhanced apoptosis as well as sensitivity of OC cells to cisplatin. In vivo results further confirmed the inhibitory effect of CDCA3 knockdown on tumor growth. Our findings indicated that CDCA3 may play an important role in OC progression, and may serve as a potential therapeutic target for the OC treatment.

Expression of DDB1 is associated with subtypes of epithelial ovarian cancer and predicts clinical outcomes

Ovarian cancer is the most lethal gynaecological malignancy. Damage specific DNA-binding protein 1 (DDB1) functions in nucleotide-excision repair and has been reported to be involved in cancer development. In this study, we aimed to determine the expression levels of DDB1 and their association with the clinical outcomes of patients with ovarian cancer. Tissue arrays were performed on 54 epithelial ovarian cancer (EOC) samples. Immunohistochemistry was performed to determine DDB1 expression. DDB1 expression levels among different EOC subtypes were analysed via one-way analysis of variance using SPSS Statistics 19.0. Correlation between DDB1 expression and chemotherapy course/progression-free survival (PFS) of patients was determined via Kaplan-Meier survival analysis using GraphPad Prism 5. Moreover, knockdown of DDB1 in ovarian cancer cells ES2 and OVCAR3 was used to preliminarily validate the role of DDB1. DDB1 was detected in the cytoplasm, especially in the nucleus, of all subtypes of EOC. However, DDB1 expression levels were significantly different between clear cell carcinoma and low-grade serous carcinoma (P = 0.022) and clear cell carcinoma and endometrioid cancer (P = 0.016). In addition, DDB1 expression was not significantly correlated with chemotherapy course (P = 0.433) or PFS (P = 0.566). High expression levels of DDB1 were correlated with significantly worse overall survival (P = 0.017) in patients with EOC. In addition, DDB1 knockdown in ovarian cancer cells decreased their proliferation in vitro. Our results revealed that DDB1 expression is heterogeneous in ovarian cancer, suggesting its use as a potential biomarker for poor survival in ovarian cancer.

PSME3 induces radioresistance and enhances aerobic glycolysis in cervical cancer by regulating PARP1

Cervical cancer (CC) ranks the fourth in gynecologic cancers. The incidence and mortality of CC has been decreased due to the cancer screening and early treatments in recent years, but the prognosis of CC patients at advanced stage is still sorrowful. Whether PSME3 exerted a role in the radioresistance of CC cells remains to be investigated. In this study, the expression of PSME3 in mRNA and protein levels was measured by RT-qPCR and western blot analysis, and increased expression of PSME3 in CC tissues and cells was observed. CCK-8 and colony formation assay revealed that the cell viability and proliferation of Hela and CaSki cells treated with different doses of X-ray was reduced due to the depletion of PSME3, indicating that silencing of PSME3 enhanced the radiosensitivity of CC cells. In addition, repair on DNA damage in CC cells was enhanced by PSME3 and the damage was attenuated by PSME3. Besides, the expression of glycolysis-related proteins (GLUT1, PGC-1α, LDHA and HK2) were enhanced by PSME3 but reduced by silencing PSME3 in CC cells. PSME3 restraint attenuated the levels of glucose consumption and lactate production, suggesting PSME3 depletion suppressed abnormal glycolysis of CC cells. Mechanically, PSME3 increased the PARP1 expression via elevating c-myc. Finally, we observed PSME3 attenuation inhibited CC growth in vivo. In conclusion, PSME3 enhanced radioresistance and aerobic glycolysis in CC by regulating PARP1, which might shed a light into the function of PSME3 in CC treatment.

DLX4 promotes the expression of PD-L1 through GATA1 in Gestational Trophoblastic Neoplasia

Gestational Trophoblastic Neoplasia (GTN) is a highly malignant tumor that originates from trophoblastic cells during embryonic development. In this study, we observed that DLX4, a member of the Distal-Less Homeobox (Dlx) gene family, was upregulated in GTN tissues and cell lines. Bioinformatic analysis showed that DLX4 was highly expressed in most cancers and had a poor survival prognosis in certain tumors; further analysis showed that DLX4 was significantly associated with genes of immune pathways and immune infiltration. Functional analyses revealed that DLX4 overexpression or knockdown did not affect GTN cell proliferation; however, we observed that DLX4 could regulate PD-L1 expression via GATA1. The luciferase reporter activity of the wild-type construct increased after overexpression of GATA1, whereas the mutation of the binding sites abolished the transcriptional increase. In conclusion, our findings suggest that DLX4 regulates PD-L1 expression via GATA1 in GTN and may be a new target for antitumor therapy.

A comprehensive study on the multi-class cervical cancer diagnostic prediction on pap smear images using a fusion-based decision from ensemble deep convolutional neural network

The diagnosis of cervical dysplasia, carcinoma in situ and confirmed carcinoma cases is more easily perceived by commercially available and current research-based decision support systems when the scenario of pathologists to patient ratio is small. The treatment modalities for such diagnosis rely exclusively on precise identification of dysplasia stages as followed by The Bethesda System. The classification based on The Bethesda System is a multiclass problem, which is highly relevant and vital. Reliance on image interpretation, when done manually, introduces inter-observer variability and makes the microscope observation tedious and time-consuming. Taking this into account, a computer-assisted screening system built on deep learning can significantly assist pathologists to screen with correct predictions at a faster rate. The current study explores six different deep convolutional neural networks- Alexnet, Vggnet (vgg-16 and vgg-19), Resnet (resnet-50 and resnet-101) and Googlenet architectures for multi-class (four-class) diagnosis of cervical pre-cancerous as well as cancer lesions and incorporates their relative assessment. The study highlights the addition of an ensemble classifier with three of the best deep learning models for yielding a high accuracy multi-class classification. All six deep models including ensemble classifier were trained and validated on a hospital-based pap smear dataset collected through both conventional and liquid-based cytology methods along with the benchmark Herlev dataset.

Substrate stiffness affects the morphology, proliferation, and radiosensitivity of cervical squamous carcinoma cells

Cervical cancer is associated with the highest morbidity rate among gynecological cancers. Radiotherapy plays an important role in the treatment of cervical cancer. However, a considerable number of patients are radiation resistant, leading to a poor prognosis. Matrix stiffness is related to the occurrence, development, and chemoresistance of solid tumors. The association between matrix stiffness and radiosensitivity in cervical cancer cells remains unknown. Here, we sought to determine the effect of matrix stiffness on the phenotype and radiosensitivity of cervical cancer cells. Cervical squamous carcinoma SiHa cells were grown on substrates of different stiffnesses (0.5, 5, and 25 kPa). Cell morphology, proliferation, and radiosensitivity were examined. Cells grown on hard substrates displayed stronger proliferative activity, larger size, and higher differentiation degree, which was reflected in a more mature skeleton assembly, more abundant pseudopodia formation, and smaller nuclear/cytoplasmic ratio. In addition, SiHa cells exhibited stiffness-dependent resistance to radiation, possibly via altered apoptosis-related protein expression. Our findings demonstrate that matrix stiffness affects the morphology, proliferation, and radiosensitivity of SiHa cells. Tissue stiffness may be an indicator of the sensitivity of a patient to radiotherapy. Thus, the data provide insights into the diagnosis of cervical cancer and the design of future radiotherapies.

RNF144B-mediated p21 degradation regulated by HDAC3 contribute to enhancing ovarian cancer growth and metastasis

We have shown before that HDAC3 was involved in the pathogenesis of ovarian cancer; however, the specific mechanism of HDAC3 on the pathogenesis of ovarian cancer has not been thoroughly studied. To explore the related proteins in the mechanism of HDAC3 on ovarian cancer. The transcriptome profiles were identified in ovarian carcinoma cells with HDAC3 knockdown or overexpression. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis were used to verify transfection efficiency. Immunofluorescence staining were performed to detect the expression levels of HDAC3 and RNF144B in tissues. Cell proliferation, apoptosis, migration and invasion were confirmed by cell counting Kit-8 (CCK-8), terminal deoxynucleotidyl transferase (TUNEL) and transwell assay, respectively. The protein expression of p53, p21, Bax and Bcl-2 was confirmed by western blot, and CoIP assay was used to validate RNF144B/P21/P53 interaction. Meanwhile, the protein synthesis inhibitor cycloheximide (CHX) was performed to treat cells to probe p21 stability. Finally, we established an in vivo tumor model to explore the effects of HDAC3 and RNF144B on tumor growth. Microarray results showed that among the overlapping genes in the two profiles (HDAC3 knockdown and overexpression), RNF144B was decreased or increased in ovarian carcinoma cells with HDAC3 knockdown or overexpression, HDAC3 overexpression promoted RNF144B expression, and HDAC3 knockdown hindered RNF144B levels. The levels of HDAC3 and RNF144B in malignant ovarian cancer were significantly higher than those in normal ovarian tissue and benign ovarian cancer tissue. RNF144B promoted cell proliferation, migration and invasion, and inhibited cell apoptosis. In addition, overexpression of HDAC3 or RNF144B inhibited p53/p21/Bax expression and promoted Bcl-2 expression. Knockout of HDAC3 or RNF144B has the opposite effect, and RNF144B interacted with p21 and regulated the p21/p53 complex degradation, and finally in vivo experiments proved that HDAC3 and RNF144B promoted tumor growth. RNF144B-mediated p21 degradation regulated by HDAC3 contributed to enhancing ovarian cancer growth and metastasis.

Paeonol inhibits the progression of endometrial cancer by affecting TRIM26-mediated LDHA ubiquitination modification

Endometrial cancer (EC) is a common gynecological malignancy characterized by abnormal glucose metabolism. Paeonol (Pae), a natural phenolic compound derived from traditional Chinese medicine, exhibits broad-spectrum antitumor activity. However, its role in modulating glycolysis and the underlying molecular mechanisms in EC remain poorly understood. The effects on EC cell viability (CCK-8), proliferation (EdU), apoptosis (flow cytometry), invasion (transwell), migration (wound healing), and tube formation rate were assessed. Glycolytic parameters were measured using corresponding commercial kits. Protein and mRNA expression levels were determined by Western blotting and RT-qPCR. The interaction between tripartite motif protein 26 (TRIM26) and lactate dehydrogenase A (LDHA) was investigated through co-immunoprecipitation (Co-IP), cycloheximide (CHX) chase, and ubiquitination assays. A xenograft model was established to examine the in vivo efficacy of Pae. Pae inhibited proliferation, metastasis, tube formation, and glycolysis of EC cells, and induced apoptosis. Pae suppressed EC malignant behaviors by downregulating LDHA expression. TRIM26 promoted ubiquitination-mediated degradation of LDHA. Overexpression of LDHA reversed the tumor-suppressive effects of TRIM26 overexpression in EC cells. TRIM26 knockdown attenuated the antitumor effects of Pae. In vivo experiments demonstrated that Pae inhibited tumor growth and regulated TRIM26/LDHA expression. Pae was found to promote TRIM26 expression, which in turn enhanced TRIM26-mediated ubiquitination and degradation of LDHA, thereby contributing to glycolysis inhibition and suppression of EC progression. These results suggested that Pae might exert its effects by modulating the TRIM26/LDHA axis and supported its potential therapeutic value in EC.

PBX1-promoted SFRP4 transcription inhibits cell proliferation and epithelial-mesenchymal transition in endometrial carcinoma

To explore the effects and mechanisms of action of the PBX1/secreted frizzled-related protein 4 (SFRP4) axis in endometrial carcinoma (EC). The expression of PBX1 and SFRP4 was analyzed using bioinformatics prediction, followed by validation in EC cells using quantitative reverse transcription-polymerase chain reaction and western blotting. After transduction with overexpression vectors for PBX1 and SFRP4, migration, proliferation, and invasion of EC cells were measured, accompanied by the detection of E-cadherin, Snail, N-cadherin, Vimentin, β-catenin, GSK-3β, and C-myc expression. The association between PBX1 and SFRP4 was validated using dual luciferase reporter gene and chromatin immunoprecipitation assays. PBX1 and SFRP4 were downregulated in EC cells. Overexpression of PBX1 or SFRP4 resulted in weakened cell proliferation, migration, and invasion, as well as decreased expression of Snail, N-cadherin, Vimentin, β-catenin, GSK-3β, and C-myc and increased expression of E-cadherin. PBX1 bound to the SFRP4 promoter and promoted its transcription. Knockdown of SFRP4 reversed the repression of overexpressed PBX1 in the malignant phenotypes and EMT of EC cells, and PBX1 repressed Wnt/β-catenin pathway activation by upregulating SFRP4 transcription. PBX1 inhibited activation of the Wnt/β-catenin pathway by promoting SFRP4 transcription, thereby suppressing malignant phenotypes in EC cells and the EMT process.

In vitro assessment of roles of PPP1R14B in cervical and endometrial cancer

Cervical and endometrial cancers are common gynecologic cancers. Protein phosphatase 1 regulatory subunit 14B (PPP1R14B) is aberrantly expressed in several tumors, while its functions in cervical and endometrial cancers remain largely uncertain. The differentially expression of PPP1R14B in cervical and endometrial cancers was predicted by GEPIA2 and Human Protein Atlas databases. The diagnostic value was analyzed by AUC curve. The association between PPP1R14B expression and overall survival was predicted using Kaplan-Meier Plotter database. The function of PPP1R14B was investigated according to in vitro assessment. PPP1R14B and phosphorylation level of Akt were analyzed through western blotting. Cell proliferation was investigated by CCK-8 and EdU staining assays. Cell apoptosis was evaluated via TUNEL staining and caspase-3 activity assays. PPP1R14B level was upregulated in cervical and endometrial cancers, and it was associated with diagnosis and worse prognosis. PPP1R14B silencing constrained cell proliferation and promoted cell death in cervical and endometrial cancers cells. PPP1R14B knockdown suppressed activation of the Akt pathway. Re-activation of the Akt signaling reversed the anti-proliferative and cell death-promoting roles of PP1R14B knockdown in cervical and endometrial cancers cells. In conclusion, PPP1R14B knockdown represses cell proliferation and facilitates cell death by inhibiting the activation of the Akt signaling in cervical and endometrial cancers.

A two-stream decision fusion network for cervical pap-smear image classification tasks

Deep learning, especially Convolution Neural Networks (CNNs), has demonstrated superior performance in image recognition and classification tasks. They make complex pattern recognition possible by extracting image features through layers of abstraction. However, despite the excellent performance of deep learning in general image classification, its limitations are becoming apparent in specific domains such as cervical cell medical image classification. This is because although the morphology of cervical cells varies between normal, diseased and cancerous, these differences are sometimes very small and difficult to capture. To solve this problem, we propose a two-stream feature fusion model comprising a manual feature branch, a deep feature branch, and a decision fusion module. Specifically, We process cervical cells through a modified DarkNet backbone network to extract deep features. In order to enhance the learning of deep features, we have devised scale convolution blocks to substitute the original convolution, termed Basic convolution blocks. The manual feature branch comprises a range of traditional features and is linked to a multilayer perceptron. Additionally, we design three decision feature channels trained from both manual and deep features to enhance the model performance in cervical cell classification. Our proposed model demonstrates superior performance when compared to state-of-the-art cervical cell classification models. We establish a 15-category 148762 cervical cytopathology image dataset (CCID). In addition, we additionally conducted experiments on the SIPaKMeD dataset. Numerous experiments show that our proposed model performs excellently compared to state-of-the-art classification models. The outcomes illustrate that our approach can significantly aid pathologists in accurately evaluating cervical smears.