OVCAR-3 Cells Research Articles

Abstract 5797: NTX301, a novel DNA hypomethylating agent, targets lipid metabolism in ovarian cancer

Abstract Background: DNA methylation causes the silencing of tumor suppressor and differentiation-associated genes being linked to chemoresistance and stemness. Previous studies demonstrated that hypomethylating agents (HMA) re-sensitize ovarian cancer (OC) cells to chemotherapy, however, translation to the clinic has been slow. NTX 301 (PinotBio) is a novel, highly potent, and orally bioavailable HMA, in early clinical development. Methods: We assessed the anti-tumor effects of NTX301 in OC models and studied the underlying molecular mechanisms by using cell proliferation, stemness, and ferroptosis assays, RNA sequencing, and validation. Results: OC cells (SKOV3, IC50=5.089nM; OVCAR5 IC50=3.664nM) were highly sensitive to NTX301 (p<0.05) compared to immortalized fallopian tube epithelial cells (FT-190) (IC50=103.3nM). Treatment with NTX301 induced significant downregulation of the DNA methyltransferases (DNMTs) 1-3 expression in SKOV3 and OVCAR5 cells compared with decitabine (p<0.05). Treatment with low dose NTX301 (100nM) induced significant transcriptomic reprogramming with 15,000 differentially expressed genes (DEGs) compared to DMSO (p<0.05). Among the NTX301 down-regulated DEGs, Gene Ontology (GO) enrichment analysis identified pathways related to the regulation of alcohol, cholesterol, and fatty acid biosynthetic process and molecular functions related to aldehyde dehydrogenase (ALDH) and oxidoreductase activity, known features of cancer stem cells (CSCs). Indeed, treatment with low dose NTX301 (100nM) reduced the ALDH(+) cell population and expression of stemness-associated transcription factors (Oct4 and Sox2). Additionally, stearoyl-Coenzyme A desaturase 1 (SCD), a key enzyme that regulates unsaturated fatty acid homeostasis in the lipogenesis process was found among the top DEGs downregulated in response to NTX3 (fold change=9.36, FDR<0.05). As blockade of SCD had been shown to induce ferroptosis, oxidized lipids levels were measured by C11-BODIPY staining. NTX301 treatment increased the levels of oxidized lipids compared to DMSO and this was blunted by deferoxamine, indicating that NTX301 promoted cell death via ferroptosis. NTX301-induced ferroptosis was successfully rescued by the addition of oleic acid (200μM) into the culture medium, supporting that it was mediated through SCD depletion. In vivo, monotherapy with NTX301 (0.5mg/kg), significantly inhibited subcutaneous OC xenograft tumor growth (p<0.05). Decreased SCD levels and increased oxidized lipids were recorded in NTX301-treated xenografts. Conclusions: NTX301 is a potent HMA active in OC models. Our findings point to a new mechanism by which epigenetic blockade disrupts lipid homeostasis and promotes cancer cell death. Citation Format: Yinu Wang, Xiaolei Situ, Horacio Cardenas, Daniela Matei, Ellie Siu, Sayedabdulrazzaq A. Alhunayan. NTX301, a novel DNA hypomethylating agent, targets lipid metabolism in ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5797.

Abstract 2668: Long non-cording RNA TUG1 promotes cisplatin resistance in ovarian cancer via DNA polymerase eta

Abstract Ovarian cancer (OVCA) is the most malignant gynecologic cancer. Despite the use of surgery and platinum-based chemotherapy, the 5-year survival rate is approximately 30% at the time of diagnosis. Cisplatin inhibits DNA synthesis by cross-linking DNA and then induces cell apoptosis. However, cisplatin resistance often causes treatment failure and tumor relapse in OVCA patients. To overcome the cisplatin resistance in OVCAs, we studied the mechanism of chemoresistance by a long non-coding RNA (lncRNA), TUG1 (taurine upregulated gene 1), which is associated with carcinogenesis and drug sensitivity in OVCAs. The expression level of TUG1 in cisplatin-resistant OVCA clinical samples is significantly higher than that of cisplatin-sensitive samples by public database. A depletion of TUG1 increased the sensitivity of OVCA cells to cisplatin together with upregulation of a set of miRNAs in vitro. We examined the targets of miRNAs and found that TUG1 depletion downregulates DNA polymerase eta (Polη) by upregulating the miRNAs. Polη is a major translesion DNA synthesis (TLS) DNA polymerase, which bypasses DNA lesions produced by DNA damaging agents and helps DNA replication. The expression level of Polη showed a positive correlation with that of TUG1 in OVCA clinical samples. Overexpression of Polη recovered cisplatin resistance in TUG1-depleted cells. Finally, cisplatin treatment combined with TUG1 depletion effectively inhibited tumor growth in xenograft mouse models, indicating the efficacy of TUG1 depletion in vivo. Our data provide a strong rationale for targeting TUG1 as a novel therapeutic approach to overcome cisplatin resistance in OVCA. Citation Format: Ryosuke Sonobe. Long non-cording RNA TUG1 promotes cisplatin resistance in ovarian cancer via DNA polymerase eta [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2668.

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

BackgroundThe aim of our study was to investigate the roles and the underlying mechanisms of orthodenticle homolog 1 (OTX1) in ovarian cancer. MethodsOTX1 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). ResultsOTX1 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. ConclusionsOTX1 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.

Next Generation of Ovarian Cancer Detection Using Aptamers

Ovarian cancer is among the seven most common types of cancer in women, being the most fatal gynecological tumor, due to the difficulty of detection in early stages. Aptamers are important tools to improve tumor diagnosis through the recognition of specific molecules produced by tumors. Here, aptamers and their potential targets in ovarian cancer cells were analyzed by in silico approaches. Specific aptamers were selected by the Cell-SELEX method using Caov-3 and OvCar-3 cells. The five most frequent aptamers obtained from the last round of selection were computationally modeled. The potential targets for those aptamers in cells were proposed by analyzing proteomic data available for the Caov-3 and OvCar-3 cell lines. Overexpressed proteins for each cell were characterized as to their three-dimensional model, cell location, and electrostatic potential. As a result, four specific aptamers for ovarian tumors were selected: AptaC2, AptaC4, AptaO1, and AptaO2. Potential targets were identified for each aptamer through Molecular Docking, and the best complexes were AptaC2-FXYD3, AptaC4-ALPP, AptaO1-TSPAN15, and AptaO2-TSPAN15. In addition, AptaC2 and AptaO1 could detect different stages and subtypes of ovarian cancer tissue samples. The application of this technology makes it possible to propose new molecular biomarkers for the differential diagnosis of epithelial ovarian cancer.

Cancer-associated mesothelial cell-derived ANGPTL4 and STC1 promote the early steps of ovarian cancer metastasis.

Ovarian cancer (OvCa) preferentially metastasizes in association with mesothelial cell-lined surfaces. We sought to determine if mesothelial cells are required for OvCa metastasis and detect alterations in mesothelial cell gene expression and cytokine secretion upon interaction with OvCa cells. Using omental samples from patients with high-grade serous OvCa and mouse models with Wt1-driven GFP-expressing mesothelial cells, we validated the intratumoral localization of mesothelial cells during human and mouse OvCa omental metastasis. Removing mesothelial cells ex vivo from human and mouse omenta or in vivo using diphtheria toxin-mediated ablation in Msln-Cre mice significantly inhibited OvCa cell adhesion and colonization. Human ascites induced angiopoietin-like 4 (ANGPTL4) and stanniocalcin 1 (STC1) expression and secretion by mesothelial cells. Inhibition of STC1 or ANGPTL4 via RNAi obstructed OvCa cell-induced mesothelial cell to mesenchymal transition while inhibition of ANGPTL4 alone obstructed OvCa cell-induced mesothelial cell migration and glycolysis. Inhibition of mesothelial cell ANGPTL4 secretion via RNAi prevented mesothelial cell-induced monocyte migration, endothelial cell vessel formation, and OvCa cell adhesion, migration, and proliferation. In contrast, inhibition of mesothelial cell STC1 secretion via RNAi prevented mesothelial cell-induced endothelial cell vessel formation and OvCa cell adhesion, migration, proliferation, and invasion. Additionally, blocking ANPTL4 function with Abs reduced the ex vivo colonization of 3 different OvCa cell lines on human omental tissue explants and in vivo colonization of ID8p53-/-Brca2-/- cells on mouse omenta. These findings indicate that mesothelial cells are important to the initial stages of OvCa metastasis and that the crosstalk between mesothelial cells and the tumor microenvironment promotes OvCa metastasis through the secretion of ANGPTL4.

Heracleum persicum Desf. ex Fisch., C.A.Mey. & Avé-Lall. fruit essential oil: content, antimicrobial activity and cytotoxicity against ovarian cancer cell line

BackgroundOne of the most important aromatic species of the Apiaceae family is Heracleum persicum Desf. ex Fisch., C.A.Mey. & Avé-Lall, which used as a spice and condiment in food. It is widely used in traditional Iranian medicine due to its anti-toxic properties. The aim of this study was to evaluate the essential oil of this plant in terms of content and ingredients, cytotoxicity and antimicrobial activity.MethodsThe fruit of H. persicum was collected in June 2019 from Maragheh region of Kashan, Iran. The essential oil was extracted by water distillation using Clevenger for 3 h. Identification analysis of H. persicum fruit essential oil (HPFEO) components was performed using gas chromatography-mass spectrometry (GC–MS). Evaluation of the effect of the HPFEO on the growth and proliferation of ovarian cancer cell line (OVCAR-3) was performed using MTT colorimetric method. Its antimicrobial activity was evaluated by agar diffusion method, minimum inhibitory concentration (MIC) and minimum bactericidal/fungal lethality concentration (MBC/MFC).ResultsThe results of analysis of the HPFEO by GC–MS showed that there were 35 compounds with 99.54% relative content. Hexyl butyrate (35.24%), octyl 2-methylbutyrate (11.65%), octyl isobutyrate (9.23%), and octyl acetate (8.42%) were the predominant compounds. Examination of cell survival showed that the viability of cells decreased depending on the concentration of the HPFEO in 24 h and had value of IC50 ~ 12.08 μg/ml against OVCAR-3 cell line. The strongest inhibitory activity of the HPFEO was against Gram-negative bacteria Pseudomonas aeruginosa and Salmonella paratyphi-A serotype (MIC < 62.50 μg/mL). Also, the strong inhibitory and lethal activity of this essential oil against Candida albicans (MIC and MBC 250 μg/mL was one degree weaker compared to nystatin (MIC = 125 μg/mL).ConclusionsThus, HPFEO, in addition to its traditional use, may have a strong and potential potential for the production of anti-proliferative and antimicrobial drugs.

An environmentally safe approach for the facile synthesis of anti-mutagenic fluorescent quantum dots: Property investigation and the development of novel antimicrobial applications

Quantum dots as nanocrystals of semiconductor materials are synthesized with different methods. The green synthesis of fluorescent quantum dots using natural stabilizing agents has attracted much attention. This research aims to study the ability of Teucrium polium L. extract to green synthesize fluorescent silver quantum dots (AgQDs). The green synthesis of AgQDs was proved using UV–Visible spectrophotometer, Fourier Transform Infrared spectroscopy (FTIR), X-Ray Diffraction (XRD), Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM), Energy-Dispersive X-ray spectroscopy (EDX), Photoluminescence (PL) analysis, and Dynamic Light Scattering (DLS).According to the findings, the shape of fluorescent AgQDs was spherical, and the maximum abundance of particle size distribution was between 3 and 5 nm.Thereupon, antimicrobial, anti-mutagenicity, anticancer, and antioxidant activities were evaluated to distinguish the biological features of AgQDs. Gram-positive bacteria (S. epidermidis, MIC: 31.25 μg/ml) and fungi (C. albicans, MIC: 31.25 μg/ml) were the most susceptible to AgQDs. Moreover, the AgQDs solution had higher antimicrobial activity than AgNO3. The Ames test demonstrated that AgQDs have considerable anti-mutagenic activities (% mutagenicity inhibition greater than 40) and are not mutagenic. The anticancer activity of AgQDs was distinguished using MTT and brine shrimp lethality (BSL) assays. The analysis indicated that AgQDs had a strong potential for cytotoxicity (BSL: LC50 = 2.4 μg/ml, MTT: IC50 = 0.8 μg/ml). Further, AgQDs were used for bioimaging human ovarian OVCAR3 cells, and their biocompatibility was remarkable. In the DPPH assay, AgQDs displayed high antioxidant activity (89.9% inhibition). Using AgQDs to coat materials to develop antibacterial agents revealed significant activity against C. albicans and S. epidermidis strains. Our results indicate that T. polium is a promising source for synthesizing AgQDs. Due to the potent bioactivities of AgQDs, they can be used in medicine and industry.

VEGFR3 suppression through miR-1236 inhibits proliferation and induces apoptosis in ovarian cancer via ERK1/2 and AKT signaling pathways.

Vascular endothelial growth factor receptor 3 (VEGFR3) is expressed in cancer cell lines and exerts a critical role in cancer progression. However, the signaling pathways of VEGFR3 in ovarian cancer cell proliferation remain unclear. This study aimed to demonstrate the signaling pathways of VEGFR3 through the upregulated expression of miR-1236 in ovarian cancer cells. We found that the messenger RNA and protein of VEGFR3 were expressed in the ovarian cancer cell lines, but downregulated after microRNA-1236 (miR-1236) transfection. The inhibition of VEGFR3, using miR-1236, significantly reduced cell proliferation, clonogenic survival, migration, and invasion ability in SKOV3 and OVCAR3 cells (p < 0.01). The flow cytometry results indicated that the rate of apoptotic cells in SKOV3 (38.65%) and OVCAR3 (41.95%) cells increased following VEGFR3 inhibition. Moreover, VEGFR3 stimulation (using a specific ligand, VEGF-CS) significantly increased extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (AKT) phosphorylation (p < 0.01), whereas VEGFR3 suppression reduced p-ERK1/2 (67.94% in SKOV3 and 93.52% in OVCAR3) and p-AKT (59.56% in SKOV3 and 78.73% in OVCAR3) compared to the VEGF-CS treated group. This finding demonstrated that miR-1236 may act as an endogenous regulator of ERK1/2 and AKT signaling by blocking the upstream regulator of VEGFR3. Overall, we demonstrated the important role of the miR-1236/VEGFR3 axis in ovarian cancer cell proliferation by regulating the ERK1/2 and AKT signaling that might be an effective strategy against ovarian cancer.

Cytotoxicity Evaluation of Unmodified Paddlewheel Dirhodium(II,II)-Acetate/-Formamidinate Complexes and Their Axially Modified Low-Valent Metallodendrimers.

Two diphenyl formamidine ligands, four dirhodium(II,II) complexes, and three axially modified low-valent dirhodium(II,II) metallodendrimers were synthesized and evaluated as anticancer agents against the A2780, A2780cis, and OVCAR-3 human ovarian cancer cell lines. The dirhodium(II,II) complexes show moderate cytotoxic activity in the tested tumor cell lines, with acetate and methyl-substituted formamidinate compounds displaying increased cytotoxicity that is relative to cisplatin in the A2780cis cisplatin resistant cell line. Additionally, methyl- and fluoro-substituted formamidinate complexes showed comparable and increased cytotoxic activity in the OVCAR-3 cell line when compared to cisplatin. The low-valent metallodendrimers show some activity, but a general decrease in cytotoxicity was observed when compared to the precursor complexes in all but one case, which is where the more active acetate-derived metallodendrimer showed a lower IC50 value in the OVCAR-3 cell line in comparison with the dirhodium(II,II) tetraacetate.

Evodiamine inhibits malignant progression of ovarian cancer cells by regulating lncRNA-NEAT1/miR-152-3p/CDK19 axis.

Evodiamine (EVO) has been demonstrated to promote apoptosis of ovarian cancer cells, and upregulate miR-152-3p level in colorectal cancer. Here, we explore part of the network mechanism of EVO and miR-152-3p in ovarian cancer. The bioinformatics website, dual luciferase reporter assay, and quantitative real-time polymerase chain reaction were applied to analyze the network among EVO, lncRNA, miR-152-3p, and mRNA. The effect and mechanism of EVO on ovarian cancer cells were determined using cell counting kit-8, flow cytometry, TUNEL, Western blot, and rescue experiments. As a result, EVO dose-dependently attenuated cell viability, induced G2/M phase arrest and apoptosis, promoted miR-152-3p level (4.5- or 2-fold changes), and inhibited expressions of NEAT1 (0.225- or 0.367-fold changes), CDK8 (0.625- or 0.571-fold changes), and CDK19 (0.25- or 0.147-fold changes) in OVCAR-3 and SKOV-3 cells. In addition, EVO decreased Bcl-2 expression, but increased the expressions of Bax and c-caspase-3. NEAT1 targeted miR-152-3p which bound to CDK19. The impacts of EVO on cell viability, cycle, apoptosis, and apoptosis-related proteins were partially reversed by miR-152-3p inhibitor, NEAT1 overexpression, or CDK19 overexpression. Furthermore, miR-152-3p mimic offset the effects of NEAT1 or CDK19 overexpression. The role of NEAT1 overexpression in the biological phenotype of ovarian cancer cells was counteracted by shCDK19. In conclusion, EVO attenuates ovarian cancer cell progression via the NEAT1-miR-152-3p-CDK19 axis.

Evaluation of Biological Activities of Twenty Flavones and In Silico Docking Study

This work aimed to evaluate the biological activities of 20 flavones (M1 to M20) and discuss their structure-activity relationships. In vitro assays were established to assess their numerous biological activities (anti-α-amylase, anti-acetylcholinesterase, anti-xanthine oxidase, anti-superoxide dismutase, and anticancer cell lines (HCT-116, MCF7, OVCAR-3, IGROV-1, and SKOV-3 cells lines)). An in silico docking study was also established in order to find the relationship between the chemical structure and the biological activities. In vitro tests revealed that M5 and M13 were the most active in terms of anti-α-amylase activity (IC50 = 1.2 and 1.4 µM, respectively). M17 was an inhibitor of xanthine oxidase (XOD) and performed better than the reference (allopurinol), at IC50 = 0.9 µM. M7 presented interesting anti-inflammatory (IC50 = 38.5 µM), anti-supriode dismutase (anti-SOD) (IC50 = 31.5 µM), and anti-acetylcholinesterase (IC50 = 10.2 µM) activities. Those abilities were in concordance with its high scavenging activity in antioxidant ABTS and DPPH assays, at IC50 = 6.3 and 5.2 µM, respectively. Selectivity was detected regarding cytotoxic activity for those flavones. M1 (IC50 = 35.9 µM) was a specific inhibitor to the MCF7 cancer cell lines. M3 (IC50 = 44.7 µM) and M15 (IC50 = 45.6 µM) were particularly potent for the OVCAR-3 cell line. M14 (IC50 = 4.6 µM) contributed more clearly to inhibiting the colon cancer cell line (HCT116). M7 (IC50 = 15.6 µM) was especially active against the ovarian SKOV human cancer cell line. The results of the biological activities were supported by means of in silico molecular docking calculations. This investigation analyzed the contribution of the structure-activity of natural flavones in terms of their biological properties, which is important for their future application against diseases.

Overexpression of CLDN16 in ovarian cancer is modulated by PI3K and PKC pathways

Epithelial ovarian cancer (EOC) is the gynecological malignant tumor of poorest prognosis and higher mortality rate. Chemotherapy is the base of high-grade serous ovarian cancer (HGSOC) treatment; however, it favors the emergence of chemoresistance and metastasis. Thus, there is an urge to search for new therapeutic targets, such as proteins related to cellular proliferation and invasion. Herein, we investigated the expression profile of claudin-16 (CLDN16 protein and CLDN16 transcript) and its possible functions in EOC. In silico analysis of CLDN16 expression profile was performed using data extracted from GENT2 and GEPIA2 platforms. A retrospective study was carried out with 55 patients to evaluate the expression of CLDN16. The samples were evaluated by immunohistochemistry, immunofluorescence, qRT-PCR, molecular docking, sequencing, and immunoblotting assays. Statistical analyzes were performed using Kaplan-Meier curves, one-way ANOVA, Turkey posttest. Data were analyzed using GraphPad Prism 8.0. In silico experiments showed that CLDN16 is overexpressed in EOC. 80.0% of all EOC types overexpressed CLDN16, of which in 87% of the cases the protein is restricted to cellular cytoplasm. CLDN16 expression was not related to tumor stage, tumor cells differentiation status, tumor responsiveness to cisplatin, or patients' survival rate. When compared to data obtained from in silico analysis regarding EOC stage and degree of differentiation, differences were found in the former but not in the later, neither in survival curves. CLDN16 expression in HGSOC OVCAR-3 cells increased by 1.95-fold (p < 0.001), 2.32-fold (p < 0.001), and 6.57-fold (p < 0.001) via PKC, PI3K, and estrogen pathways, respectively. Altogether, our results suggest that despite the low number of samples included in our in vitro studies, adding to the expression profile findings, we provided a comprehensive study of CLDN16 expression in EOC. Therefore, we hypothesize that CLDN16 is a potential target in the diagnosis and treatment of the disease.

The inhibitory effect of 6-gingerol and cisplatin on ovarian cancer and antitumor activity: In silico, in vitro, and in vivo.

Epithelial ovarian cancer is very common in women and causes hundreds of deaths per year worldwide. Chemotherapy drugs including cisplatin have adverse effects on patients' health. Complementary treatments and the use of herbal medicines can help improve the performance of medicine. 6-Gingerol is the major pharmacologically active component of ginger. In this study, we compared the effects of 6-gingerol, cisplatin, and their combination in apoptotic and angiogenetic activities in silico, in test tubes, and in in vivo assays against two ovarian cancer cell lines: OVCAR-3 and human umbilical vein endothelial cells (HUVECs). The drug-treated cell lines were evaluated for their cytotoxicity, cell cycle, and apoptotic and angiogenetic gene expression changes. The proportion of apoptosis treated by 6-gingerol coupled with cisplatin was significantly high. In the evaluation of the cell cycle, the combination therapy also showed a significant promotion of a higher extent of the S sequence. The expression of p53 level, Caspase-8, Bax, and Apaf1 genes was amplified again with combination therapy. Conversely, in both cell lines, the cumulative drug concentrations reduced the expression of VEGF, FLT1, KDR, and Bcl-2 genes. Similarly, in the control group, combination treatment significantly decreased the expression of VEGF, FLT1, KDR, and Bcl-2 genes in comparison to cisplatin alone. The findings of the present study demonstrated that the cisplatin and 6-gingerol combination is more effective in inducing apoptosis and suppressing the angiogenesis of ovarian cancer cells than using each drug alone.

Study on the effect of 5-aminolevulinic acid-mediated photodynamic therapy combined with cisplatin on human ovarian cancer OVCAR-3 ​cells

PurposeThis article explores the effect of 5-aminolevulinic acid (5-ALA)-mediated photodynamic therapy (PDT) combined with cisplatin (CDDP) on the apoptosis of human ovarian cancer cells and the mechanism of action of the combination therapy. Materials and methodsHuman ovarian cancer OVCAR-3 ​cells were cultured in vitro and divided into 5-ALA/PDT group, CDDP group and combined treatment group (5-ALA/PDT combined with different concentrations of CDDP). After administration of the corresponding drugs, a CCK-8 assay was used to detect the inhibition rate of cell proliferation. After Rhodamine 123 staining, mitochondrial membrane potential changes were observed under fluorescence microscopy. The apoptosis rate and reactive oxygen species (ROS) content were detected by flow cytometry. Western blotting was used to detect protein expression. ResultsThe CCK-8 assay showed that CDDP in combination with 5-ALA/PDT significantly enhanced cytotoxicity compared to treatment with CDDP alone and that low doses of CDDP were sufficient to induce these combination effects. The mitochondrial membrane potential in each combination treatment group gradually decreased with increasing CDDP concentration, while the apoptosis rate and reactive oxygen species (ROS) content detected by flow cytometry gradually increased. Western blotting assay showed that the expression of bax, cleaved caspase-9, cleaved caspase-3, and cleaved PARP was increased, while the expression of bcl-2, caspase-9, caspase-3, and PARP was decreased, and the differences were statistically significant (P ​< ​0.05). ConclusionsIn summary, 5-ALA/PDT combined with CDDP can effectively inhibit cell proliferation and promote apoptosis, and this combination may induce apoptosis by activating the mitochondrial pathway.

Elevated Levels of Lamin A Promote HR and NHEJ-Mediated Repair Mechanisms in High-Grade Ovarian Serous Carcinoma Cell Line

Extensive research for the last two decades has significantly contributed to understanding the roles of lamins in the maintenance of nuclear architecture and genome organization which is drastically modified in neoplasia. It must be emphasized that alteration in lamin A/C expression and distribution is a consistent event during tumorigenesis of almost all tissues of human bodies. One of the important signatures of a cancer cell is its inability to repair DNA damage which befalls several genomic events that transform the cells to be sensitive to chemotherapeutic agents. This genomic and chromosomal instability is the most common feature found in cases of high-grade ovarian serous carcinoma. Here, we report elevated levels of lamins in OVCAR3 cells (high-grade ovarian serous carcinoma cell line) in comparison to IOSE (immortalised ovarian surface epithelial cells) and, consequently, altered damage repair machinery in OVCAR3. We have analysed the changes in global gene expression as a sequel to DNA damage induced by etoposide in ovarian carcinoma where lamin A is particularly elevated in expression and reported some differentially expressed genes associated with pathways conferring cellular proliferation and chemoresistance. We hereby establish the role of elevated lamin A in neoplastic transformation in the context of high-grade ovarian serous cancer through a combination of HR and NHEJ mechanisms.

Anticancer and Biological Effects of Some Natural Compounds and Theoretical Investigation of them Against RdRP of SARS-COV-2: In Silico and In Vitro Studies.

In this study, Skullcapflavone I and Skullcapflavone II molecules showed good inhibitory activities against α-glucosidase and sorbitol dehydrogenase enzymes with IC50 values of 102.66 ± 8.43 and 95.04 ± 11.52nM for α-glucosidase and 38.42 ± 3.82 and 28.81 ± 3.26µM for sorbitol dehydrogenase. The chemical activities of Skullcapflavone I and Skullcapflavone II against α-glucosidase and sorbitol dehydrogenase were assessed by conducting the molecular docking study. The anticancer activities of the compounds were examined against SW-626, SK-OV-3, OVCAR3, and Caov-3 cell lines. The chemical activities of Skullcapflavone I and Skullcapflavone II against some of the expressed surface receptor proteins (estrogen receptor, EGFR, androgen receptor, and GnRH receptor) in the mentioned cell lines were investigated using in silico calculations. Moreover, the activity of the compounds against RNA polymerase of SARS-COVE-2 was also assessed using the molecular modeling study. These compounds created strong contacts with the enzymes and receptors. The considerable binding affinity of the compounds to the enzymes and proteins showed their ability as inhibitors. Furthermore, even at modest dosages, these substances markedly reduced the viability of ovarian cancer cells. Additionally, the viability of ovarian cancer cells was significantly decreased by a 300 μM dosage of all compounds. Antiovarian cancer results of Skullcapflavone I on SK-OV-3, SW-626, OVCAR3, and Caov-3 were 63.14, 1.55, 19.42, and 52.04µM, respectively. Also, cytotoxicity results of Skullcapflavone II on SK-OV-3, SW-626, OVCAR3, and Caov-3 were 5.18, 21.44, 33.87, and 72.66µM, respectively.

UBE2E2 enhances Snail-mediated epithelial-mesenchymal transition and Nrf2-mediated antioxidant activity in ovarian cancer

Dissemination of ovarian cancer (OvCa) cells can lead to inoperable metastatic lesions in the bowel and omentum, which have a poor prognosis despite surgical and chemotherapeutical options. A better understanding of the mechanisms underlying metastasis is urgently needed. In this study, bioinformatics analyses revealed that UBE2E2, a less-studied ubiquitin (Ub)-conjugating enzyme (E2), was upregulated in OvCa and was associated with poor prognosis. Subsequently, we performed western blot analysis and IHC staining with 88 OvCa and 26 normal ovarian tissue samples, which further confirmed that UBE2E2 protein is highly expressed in OvCa tissue but weakly expressed in normal tissue. Furthermore, the silencing of UBE2E2 blocked OvCa cell migration, epithelial-mesenchymal transition (EMT) and metastasis in vitro, whereas UBE2E2 overexpression exerted the opposite effects. Mechanistically, UBE2E2 promoted p62 accumulation and increased the activity of the Nrf2-antioxidant response element (ARE) system, which ultimately activated the Snail signaling pathway by inhibiting the ubiquitin-mediated degradation of Snail. Additionally, co-IP and immunofluorescence demonstrated that a direct interaction exists between UBE2E2 and Nrf2, and the N-terminal of UBE2E2 (residues 1-52) is required and sufficient for its interaction with Nrf2 protein. Mutations in the active site cysteine (Cys139) impaired both the function and cellular distribution of UBE2E2. More importantly, the deletion of UBE2E2 reduced tumorigenicity and metastasis in xenograft OvCa mouse models. Taken together, our findings reveal the role of the UBE2E2-Nrf2-p62-Snail signaling axis in OvCa and thus provides novel therapeutic targets for the prevention of OvCa metastasis.

Dual regulation of Akt and glutathione caused by isoalantolactone effectively triggers human ovarian cancer cell apoptosis.

Ovarian cancer is one of leading causes of cancer death in gynecological tumor. Isoalantolactone (IL), present in several medicinal plants, exhibits various biological activities, and its mechanism underlying anti-ovarian cancer activity needs to be further investigated. Here, we find that IL inhibits the proliferation of SKOV-3 and OVCAR-3 cells by causing G2/M phase arrest and inducing apoptosis. Moreover, IL decreases intracellular glutathione (GSH) level, and induces reactive oxygen species (ROS) generation in SKOV-3 cells. Furthermore, IL induces inactivation of Akt which is required for the cytotoxicity of IL. In addition, overexpression of Akt attenuates the IL-induced growth inhibition and ROS generation. GSH supplementation moderately increases the expression of phospho-Akt. Further investigation reveals that pretreatment with L-buthionine-sulfoximine (a GSH biosynthesis inhibitor) restores the Akt-mediated attenuation of growth inhibition induced by IL. Moreover, co-treatment with IL and wortmannin (an Akt pathway inhibitor) increases the growth inhibition attenuated by pretreatment with N-acetyl-L-cysteine (a precursor for GSH biosynthesis). These results indicate that inactivation of Akt and downregulation of GSH level induced by IL are related to each other. In conclusion, combined targeting Akt and GSH is an effective strategy for cancer therapy and IL can be a promising anticancer agent for further exploration.

Synergistic antitumor effect of folate-targeted Pluronic™ F-127/poly(lactic acid) polymersomes for codelivery of doxorubicin and paclitaxel.

Aim: Folate-targeted Pluronic™ F-127/poly(lactic acid) (FA-F127-PLA) polymersomes were used as codelivery carriers of doxorubicin hydrochloride (DOX) and paclitaxel (PTX) to achieve a targetedsynergistic antitumor effect. Materials & methods: The cytotoxicity of PTX/DOX polymersomes against OVCAR-3 cells was determined by methyl thiazolyl tetrazolium assay. The cellular uptake of PTX/DOX polymersomes was examined by HPLC and micro-bicinchoninic acid techniques. Results: The polymersomes showed a bilayer core-shell structure with negative charge and good dispersion. PTX1/DOX5 polymersomes with a mass ratio of PTX to DOX of 1:5 showed the best synergistic effect and the highest cellular uptake. Conclusion: FA-F127-PLA polymersomes have thegreat promise for codelivery of multiple chemotherapeutics to achieve a targeted antitumor synergistic effect.

Overexpression of MKRN2 Inhibits the Growth of Ovarian Cancer Cells

Ovarian cancer has a high mortality with low five-year survival rates. The role of the E3 ligase Makorin ring finger protein 2 (MKRN2) in ovarian cancer is unknown. This study investigated the impact of MKRN2 on the growth of ovarian cancer. MKRN2 expression in ovarian cancer tissue was analyzed by immunohistochemistry. Overexpression of MKRN2 was induced in two ovarian cancer cell lines (SKOV3 and CAOV3) by lentivirus transfection, and expression levels were verified by western blotting. Proliferation and growth were determined by CCK-8 and colony formation assays, while migration was examined using transwell assays and apoptosis by flow cytometry. Xenograft tumors of transfected SKOV3 cells were established in mice, and immunohistochemistry and TUNEL assays measured MKRN2 levels and apoptosis in tumor cells. Reduced levels of MKRN2 in cancerous tissue relative to non-cancerous ovarian tissues. Lentiviral-based MKRN2 overexpression in SKOV3 and CAOV3 cells reduced tumor-associated behavior while inducing apoptosis in vitro. In xenograft tumors, MKRN2 overexpression inhibited ovarian cancer growth and increased apoptosis in vivo. These findings imply the MKRN2 involvement in ovarian carcinogenesis and suggest its potential for treating the disease.