Long noncoding RNA NEAT1 decreases polycystic ovary syndrome progression via the modulation of the microRNA-324-3p and BRD3 axis.

Long noncoding RNA nuclear-enriched abundant transcript 1 (NEAT1) is believed to be involved in many gynecological and obstetrics disorders. Nevertheless, the role of NEAT1 in polycystic ovary syndrome (PCOS) is scarcely investigated. Our study aimed to investigate the role of NEAT1, microRNA (miR)-324-3p, and bromodomain containing 3 (BRD3) in PCOS. First, 80 women with PCOS and 80 healthy (non-PCOS) women were included, and their serum hormone levels were tested. Next, the PCOS mouse model was established by dehydroepiandrosterone injection, and then NEAT1, miR-324-3p, and BRD3 expression levels were detected in the PCOS mice. Lentivirus carrying short hairpin-NEAT1 or miR-324-3p agomir was injected into the PCOS mice to determine the change in biochemical indices and pathology. Moreover, a rescue experiment was conducted, after which, the binding relationships among NEAT1, miR-324-3p, and BRD3 were analyzed. NEAT1 and BRD3 were expressed at a high level while miR-324-3p was expressed at a low level in women with PCOS and PCOS mice. Reduced levels of NEAT1 or elevated levels of miR-324-3p mitigated metabolic disorders and alleviated ovarian pathological changes in PCOS mice. Mechanistically, NEAT1 sponged miR-324-3p and miR-324-3p targeted BRD3. In the rescue experiment, elevated miR-324-3p or reduced BRD3 level reversed the effects of the enhanced NEAT1 on metabolic disorders and ovarian pathological changes in PCOS mice. NEAT1 exacerbates metabolic disorders and ovarian pathological changes in PCOS mice by downregulating miR-324-3p and upregulating BRD3. This study gives a novel direction in PCOS treatment.

Roles of Long Non-Coding RNA CCAT2 in Cervical Cancer Cell Growth and Apoptosis.

BACKGROUND: This study aimed to analyze the potential function of lncRNA CCAT2 in cervical cancer cell proliferation and apoptosis. MATERIAL/METHODS: Expression 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. RESULTS: CCK8 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. CONCLUSIONS: LncRNA CCAT2 promoted the proliferation and survival of cervical cancer cells.

Down-regulation of Ras-related protein Rab 5C-dependent endocytosis and glycolysis in cisplatin-resistant ovarian cancer cell lines.

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, fructose-bisphosphate 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.

Key results from the salt lake regional smoke, ozone, and aerosol study (SAMOZA).

The Northern Wasatch Front area is one of ~ 50 metropolitan regions in the U.S. that do not meet the 2015 O3 standard. To better understand the causes of high O3 days in this region we conducted the Salt Lake regional Smoke, Ozone and Aerosol Study (SAMOZA) in the summer of 2022. The primary goals of SAMOZA were: Measure a suite of VOCs, by Proton Transfer Reaction Mass Spectrometry (PTR-MS) and the 2,4-dinitrophenylhydrazine (DNPH) cartridge method.Evaluate whether the standard UV O3 measurements made in SLC show a positive bias during smoke events, as has been suggested in some recent studies.Use the observations to conduct photochemical modeling and statistical/machine learning analyses to understand photochemistry on both smoke-influenced and non-smoke days.Implications: The Northern Wasatch Front area is one of ~50 metropolitan regions in the U.S. that do not meet the 2015 O3 standard. To better understand the causes of high O3 days in this region we conducted the Salt Lake regional Smoke, Ozone and Aerosol Study (SAMOZA) in the summer of 2022. A number of policy relevant findings are identified in the manuscript including role of smoke and NOx vs VOC sensitivity.

We found no significant difference in the O₃ measurements using a "scrubber-less" UV instrument compared to the standard O₃ measurements at PM_2.5 concentrations up to 60 µg m⁻³.On days with smoke, we found that PM_2.5, CO, O₃ and nearly all VOCs were significantly enhanced. On average, NO_x was also enhanced on days with smoke, but this was complicated by day of week effects.Photochemical modeling of O₃ production rates at the Utah Tech Center demonstrates a strong sensitivity to VOC concentrations and less sensitivity to NOx. For non-smoke days, achieving the current O₃ standard would require regional reductions in VOCs of ~40% or reductions in NO_x ~ 60%.The photochemical modeling shows that formaldehyde and other OVOCs, along with alkenes, were the most important O₃ precursors.Generalized Additive Modeling (GAM) gave similar MDA8 O₃ enhancements on smoky days as the photochemical modeling. Analysis of the GAM results show that 23% of the smoke days have GAM residuals that exceed the U.S. EPA's criteria for inclusion as exceptional event documentation.

Development and validation of nomogram for the prediction of preterm delivery based on patient characteristics and circulating inflammatory cells in patients with gestational diabetes mellitus.

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: χ2=3.618, P=0.306 in the training cohort; χ2=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.

Metabolomics profiling of maternal and umbilical cord blood in normoglycemia macrosomia.

Background: Macrosomia is a common disorder that occurs during pregnancy. We investigated the comprehensive metabolite profiles of pregnant maternal and fetal sera in normoglycemic macrosomia in a Chinese population. Methods: Forty pregnant women and their fetuses were included in the study (twenty macrosomia patients and twenty normal-weight controls). Maternal and umbilical cord serum metabolites were identified using ultra-performance liquid chromatography coupled with tandem mass spectrometry. Results: In total, 203 metabolites were identified. Lipids and lipid-like molecules were the predominant metabolites. Fifty-three metabolites with significant differences were obtained in the maternal samples. In the macrosomia group, the levels of docosahexaenoic acid, eicosapentaenoic acid, and arachidonic acid were significantly higher than those in the control group. Umbilical cord serum samples were obtained for 24 different metabolites. The maternal-fetal gradient of polyunsaturated fatty acids was decreased in the macrosomia group. Aconitic acid, citric acid, isocitric acid, 2-methylhexanoic acid, and 12-hydroxystearic acid were the common differential metabolites in the maternal and umbilical cord serum samples. Conclusion: There were obvious metabolic abnormalities in the sera of pregnant women and fetuses with macrosomia. Lipids and lipid-like molecules were the predominant differential metabolites but had different classifications in the maternal and umbilical cord serum. These results may provide new insights into the long-term metabolic disorders associated with macrosomia.

Molecular Mechanism of Curcumin Derivative on YAP Pathway against Ovarian Cancer.

The purpose of this study is to study the effect of curcumin derivative WZ10 on the proliferation, invasion and apoptosis of ovarian cancer OVCAR3 cells, and to explore its mechanism. MTT assay was used to detect the effect of WZ10 on the proliferation of ovarian cancer OVCAR3 cells; Annexin V/PI double staining flow cytometry was used to detect the effect of WZ10 on cell apoptosis; Transwell method was used to detect the effect of WZ10 on cell invasiveness; Western blot was used to investigate the effect of WZ10 Mechanisms affecting OVCAR3 activity in ovarian cancer in vitro. Our results show that WZ10 treatment could significantly inhibit the proliferation and invasion of OVCAR3 cells, induce apoptosis of OVCAR3 cells in a dose-dependent manner. After knockdown of Hippo expression with sh-RNA, further combined treatment with WZ10 had no significant image on ovarian cancer OVCAR3 cells. In conclusion: WZ10 can significantly inhibit the proliferation of OVCAR3 cells, reduce cell invasion and proliferation by downregulating the activation of Hippo-YAP pathway, and induce cell apoptosis.

Bisphenol A stimulates differentiation of rat stem Leydig cells in vivo and in vitro.

Bisphenol A (BPA) is widely used in consumer products and a potential endocrine disruptor linked with sexual precocity. However, its action and underlying mechanisms on male sexual maturation is unclear. In the present study, we used a unique in vivo ethane dimethane sulfonate (EDS)-induced Leydig cell regeneration model that mimics the pubertal development of Leydig cells and an in vitro stem Leydig cell differentiation model to examine the roles of BPA in Leydig cell development in rats. Intratesticular exposure to doses (100 and 1000 pmol/testis) of BPA from post-EDS day 14-28 stimulated Leydig cell developmental regeneration process by increasing serum testosterone level and Leydig cell-specific gene (Lhcgr, Star, Cyp11a1, Hsd3b1, Cyp17a1, Hsd17b3, and Hsd11b1) and their protein expression levels. BPA did not alter serum luteinizing hormone and follicle-stimulating hormone levels as well as the proliferative capacity of Leydig cells in vivo. In vitro study demonstrated that BPA (100 nmol/L) stimulated the differentiation of stem Leydig cells by increasing medium testosterone levels and up-regulating Leydig cell-specific gene (Lhcgr, Cyp11a1, Hsd3b1, Cyp17a1, and Hsd17b3) and their proteins but did not affect their proliferation measured by EdU incorporation. In conclusion, BPA stimulates the differentiation of stem Leydig cells in rat testes, thus possibly causing sexual precocity in the male.

Triclocarban and Triclosan Inhibit Human Aromatase via Different Mechanisms.

Human aromatase (CYP19A1) is an important enzyme, which produces estrogen from androgen for maintaining the female reproductive function and pregnancy. Triclocarban and triclosan are antimicrobial chemicals added to personal care, household, and industrial products. They could be endocrine disruptors and may disrupt human CYP19A1 activity. In the present study, we investigated the effects of triclocarban and triclosan on estradiol production and human CYP19A1 activity in JEG-3 cells. Triclocarban and triclosan reduced estradiol production in JEG-3 cells. Triclocarban and triclosan inhibited human CYP19A1 with IC50 values of 15.81 and 6.26 µM, respectively. Triclosan competitively inhibited CYP19A1, while triclocarban noncompetitively inhibited this enzyme. Docking study showed that triclosan bound to the steroid-binding pocket of CYP19A1, while triclocarban was off this target, suggesting a different mechanism. In conclusion, triclocarban and triclosan are inhibitors of human CYP19A1.

Phosphoproteomic analysis of gossypol-induced apoptosis in ovarian cancer cell line, HOC1a.

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.

Gossypol induces apoptosis in ovarian cancer cells through oxidative stress.

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.