BackgroundThis study aimed to analyze the potential function of lncRNA CCAT2 in cervical cancer cell proliferation and apoptosis.Material/MethodsExpression level of CCAT2 in cervical cancer cell lines (HeLa, CaSki, and SiHa) was detected by quantitative real-time PCR. CCAT2 knockdown was established by transfecting siRNA into human cervical cancer cells. Its effects on cell proliferation were studied using cell-counting kit-8 assay. The effect of CCAT2 on cervical cancer cells cycle and apoptosis was assessed by flow cytometry assay.ResultsCCK8 assay showed that CCAT2 knockdown inhibited cell proliferation in HeLa, CaSki, and SiHa cells. The flow cytometry confirmed the results that knockdown of CCAT2 could induce cervical cancer cells cycle G1 phase arrestment and trigger the cells apoptosis.ConclusionsLncRNA CCAT2 promoted the proliferation and survival of cervical cancer cells.

In the present work, metabolomic and redox proteomic analyses were carried out on an untreated- and gossypol-treated ovarian cancer cell line, SKOV3. Gossypol treatment resulted in cell death through oxidative stress. Metabolite analysis showed that gossypol induces a decrease of the cellular levels of GSH, aspartic acid, and FAD. Using a combination of double labeling and LC-MS-MS, we identified changes in thiol-redox states of 545 cysteine-containing peptides from 356 proteins. The frequently occurring amino acid residue immediately before or after the cysteine in these peptides is the non-polar and neutral leucine, valine, or alanine. These redox sensitive proteins participate in a variety of cellular processes. We have characterized the redox-sensitive cysteine residues in PKM2, HSP60, malate dehydrogenase and other proteins that play important roles in metabolism homeostasis and stress responses. The three cysteine residues of HSP60 exhibit different responses to gossypol treatment: an increase of thiol/disulfide ratio for the Cys447 residue due to a decrease of the cellular GSH level, and a decrease of thiol/disulfide ratios for Cys442 and Cys237 residues due to oxidation and sulfation. This study suggests that thiol/disulfide ratios are dependent on the level of cellular GSH. Our data provide a valuable resource for deciphering the redox regulation of proteins and for understanding gossypol-induced apoptosis in ovarian cancer cells.

Drug resistance poses a major challenge to ovarian cancer treatment. Understanding mechanisms of drug resistance is important for finding new therapeutic targets. In the present work, a cisplatin-resistant ovarian cancer cell line A2780-DR was established with a resistance index of 6.64. The cellular accumulation of cisplatin was significantly reduced in A2780-DR cells as compared with A2780 cells consistent with the general character of drug resistance. Quantitative proteomic analysis identified 340 differentially expressed proteins between A2780 and A2780-DR cells, which involve in diverse cellular processes, including metabolic process, cellular component biogenesis, cellular processes, and stress responses. Expression levels of Ras-related proteins Rab 5C and Rab 11B in A2780-DR cells were lower than those in A2780 cells as confirmed by real-time quantitative PCR and Western blotting. The short hairpin (sh)RNA-mediated knockdown of Rab 5C in A2780 cells resulted in markedly increased resistance to cisplatin whereas overexpression of Rab 5C in A2780-DR cells increases sensitivity to cisplatin, demonstrating that Rab 5C-dependent endocytosis plays an important role in cisplatin resistance. Our results also showed that expressions of glycolytic enzymes pyruvate kinase, glucose-6-phosphate isomerase, fructosebisphosphate aldolase, lactate dehydrogenase, and phosphoglycerate kinase 1 were down-regulated in drug resistant cells, indicating drug resistance in ovarian cancer is directly associated with a decrease in glycolysis. Furthermore, it was found that glutathione reductase were up-regulated in A2780-DR, whereas vimentin, HSP90, and Annexin A1 and A2 were down-regulated. Taken together, our results suggest that drug resistance in ovarian cancer cell line A2780 is caused by multifactorial traits, including the down-regulation of Rab 5C-dependent endocytosis of cisplatin, glycolytic enzymes, and vimentin, and up-regulation of antioxidant proteins, suggesting Rab 5C is a potential target for treatment of drug-resistant ovarian cancer. This constitutes a further step toward a comprehensive understanding of drug resistance in ovarian cancer.Molecular & Cellular Proteomics 13: 10.1074/ mcp.M113.033217, 3138-3151, 2014.Ovarian cancer is the major cause of death in women with gynecological cancer. Early diagnosis of ovarian cancer is difficult, while its progression is fast. The standard treatment is surgical removal followed by platinum-taxane chemotherapy. However, the efficacy of the traditional surgery and chemotherapy is rather compromised and platinum resistant cancer recurs in ϳ25% of patients within six months, and the overall five-year survival rate is about 31% (1-3). Virtually no efficient second line treatment is available. In order to increase survival rates from ovarian cancer and enhance patients' quality of life, new therapeutic targets are urgently required, necessitating a deeper understanding of molecular mechanisms of drug resistance.Mechanisms of drug-resistance in ovarian cancer have been extensi...

Ovarian cancer is a major cause for death of gynecological cancer patients. The efficacy of traditional surgery and chemotherapy is rather compromised and platinum-resistant cancer recurs. Finding new therapeutic targets is urgently needed to increase the survival rate and to improve life quality of patients with ovarian cancer. In the present work, phosphoproteomic analysis was carried out on untreated and gossypol-treated ovarian cancer cell line, HOC1a. We identified approximately 9750 phosphopeptides from 3030 phosphoproteins, which are involved in diverse cellular processes including cytoskeletal organization, RNA and nucleotide binding, and cell cycle regulation. Upon gossypol treatment, changes in phosphorylation of twenty-nine proteins including YAP1 and AKAP12 were characterized. Western blotting and qPCR analysis were used to determine expression levels of proteins in YAP1-related Hippo pathway showing that gossypol induced upregulation of LATS1, which phosphorylates YAP1 at Ser 61. Furthermore, our data showed that gossypol targets the actin cytoskeletal organization through mediating phosphorylation states of actin-binding proteins. Taken together, our data provide valuable information to understand effects of gossypol on protein phosphorylation and apoptosis of ovarian cancer cells.

Background The incidence of preterm delivery (<37 weeks’ gestation) is increased due to gestational diabetes mellitus (GDM). The preterm delivery is the leading cause of death in children. If potential preterm delivery can be diagnosed early and then prevented, adverse pregnancy outcomes can be improved. Therefore, effective methods are needed for early prediction of preterm delivery in women with GDM. Methods Patients with GDM defined as the presence of at least 1 plasma glucose abnormality at 24–28 weeks of pregnancy [fasting plasma glucose ≥5.1 mmol/L, 60-min ≥10.0 mmol/L, 120-min ≥8.5 mmol/L by 75 g oral glucose tolerance test (OGTT)] from the First Affiliated Hospital of Wenzhou Medical University were enrolled. The data (564 patients) recorded from January 2017 to June 2020 were named the training cohort, and the data (242 patients) obtained from patients with GDM, from July 2020 to January 2022, were named the validation cohort. Mann-Whitney U test and chi-square test were used to compare the skewed distributed and categorical data, respectively. According to the results of univariate logistic regression analysis, the multivariate logistic regression model was developed in the training cohort. Then, the nomogram was established. The validation of the nomogram was conducted on the training and validation cohort. Results No significant differences in baseline characteristics were detected between the 2 cohorts (all P>0.05). The multivariate analysis suggested that maternal age, insulin use, NLR, and monocyte count were the independent predictors of preterm delivery. A nomogram for predicting the probability of preterm delivery was developed. The model suggested good discrimination [areas under the curve (AUC) =0.885, 95% confidence interval (95% CI): 0.855–0.910, sensitivity =83.0%, specificity =83.1% in the training cohort; AUC =0.919, 95% CI: 0.858–0.980, sensitivity =90.6%, specificity =84.8% in the validation cohort] and good calibration [Hosmer-Lemeshow (HL) test: χ 2 =3.618, P=0.306 in the training cohort; χ 2 =6.012, P=0.111 in the validation cohort]. Conclusions The visual nomogram model appears to be a reliable approach for the prediction of preterm delivery, allowing clinicians to take timely measures to prevent the occurrence of preterm delivery in women with GDM at the time of GDM diagnosis, and deserves further investigation.