Phosphoproteomic profiling of the hippocampus of offspring rats exposed to prenatal stress.

INTRODUCTION: Prenatal stress (PS) can cause depression in offspring. However, the underlying biological mechanism of these influences is still unclear. This work was implemented to investigate the molecular mechanisms of depressive-like behavior of offspring rats insulted with PS. METHODS: Relative quantitative phosphoproteomics of the hippocampus of PS susceptibility (PS-S) and control (CON) rat offspring was performed using liquid chromatography-tandem mass spectrometry to confirm known pathways and to identify new mechanisms involved in depression. RESULTS: A total of 6790 phosphopeptides, 9817 phosphorylation sites, and 2978 phosphoproteins were detected. Among the 2978 phosphoproteins, 1760 (59.09%) had more than two phosphorylated sites, the ENSRNOP00000023460 protein had more than 117 phosphorylated sites, and the average distribution of modification sites per 100 amino acids was 2.97. There were 197 different phosphopeptides, including 140 increased phosphopeptides and 57 decreased phosphopeptides in the PS-S offspring rats, compared to the CON offspring rats. These differential phosphopeptides corresponded to 100 upregulated and 44 downregulated phosphoproteins, respectively. Gene ontology enrichment analysis revealed that these different phosphoproteins in the top five enriched terms in the cellular component, molecular function, and biological proces categories were involved in a total of 35 different phosphoproteins, and these phosphoproteins were mainly related to myelin-, microtubule- and synapse-associated proteins. The enrichment of Kyoto Encyclopedia of Genes and Genome pathways was found to be involved in many essential biological pathways, and the top five pathways included amphetamine addiction, insulin secretion, Cushing syndrome, and the circadian entrainment signaling pathway. These first five pathways were related to nine phosphoproteins, including Adcy9, Apc, Cacna1c, Camk2a, Camk2b, Camk2g, Ctnnd2, Grin2a, and Stx1a. The full data are available via ProteomeXchange with identifier PXD019117. CONCLUSION: We preliminarily identified 144 different phosphoproteins involved in myelin, microtubule, and synapse formation and plasticity in the hippocampus of susceptible offspring rats exposed to PS.

OTUB2 Promotes Homologous Recombination Repair Through Stimulating Rad51 Expression in Endometrial Cancer.

Genetic instability, raised from dysregulation of DNA repair, is involved in tumor development. OTUB2 (ovarian tumor domain protease domain-containing ubiquitin aldehyde-binding protein 2), which is responsible for DNA double-strand break (DSB), is implicated in carcinogenesis of various tumors. The effect of OTUB2 on endometrial cancer progression was then investigated. First, OTUB2 was found to be upregulated in endometrial cancer tissues and cell lines, and was closely associated with overall survival of endometrial cancer patients. Cell Counting Kit-8 and flow cytometry assay results revealed that overexpression of OTUB2 enhanced cell viability of endometrial cancer cells, while knockdown of OTUB2 inhibited cell viability. Moreover, as demonstrated by promoting cell viability and suppression of cell apoptosis, cisplatin-induced cell damage was reversed by OTUB2. Mechanistically, OTUB2 could activate Yes-associated protein/transcriptional co-activator with PDZ-binding motif (TAZ) to promote homologous recombination repair via depletion of γH2AX (phosphorylation of histone H2AX) and accumulation of Rad51. In vivo xenograft model also showed that silence of OTUB2 suppressed the growth of endometrial cancer and increased tumor sensitivity to antitumor drugs. In conclusion, OTUB2 promoted homologous recombination repair in endometrial cancer via YAP/TAZ-mediated Rad51 expression, providing a potential therapeutic target for endometrial cancer.

Long noncoding RNA TUSC7 inhibits cell proliferation, migration and invasion by regulating SOCS4 (SOCS5) expression through targeting miR-616 in endometrial carcinoma.

BACKGROUND: Long non-coding RNA (lncRNA) is emerging as an important regulator in various physiological and pathological processes. Recently, it was found that lncRNA long non-coding RNA tumor suppressor candidate 7 (TUSC7) could play tumor suppressive roles in several cancers. However, the function and underlying regulatory mechanism of lncRNA TUSC7 in endometrial carcinoma (EC) remains largely unclear. METHODS: The expression levels of TUSC7 and microRNAs-616 (miR-616) were analyzed by real-time PCR and in situ hybridization. Cell cycle and cell metastasis associated protein expressions were determined by western blotting. Cell proliferation, cycle and metastasis were determined by CCK-8 cell viability, colony formation, flow cytometer, wound scratch and transwell assays respectively in vitro. RNA pull-down, luciferase and western blotting assays were used to examine the target relationship between TUSC7 and miR-616 or that between miR-616 and suppressors of cytokine signaling 4 (5) (SOCS4 (SOCS5)). The functional effects of TUSC7 through sponging miR-616 were further examined using a xenograft tumor mouse model in vivo. RESULTS: TUSC7 was downexpressed in EC tissues and cell lines, and TUSC7 upregulation could remarkably inhibit cell proliferation, cycle progression and metastasis in EC cells. Mechanistic investigations demonstrated that TUSC7 can interact with miR-616 and decrease its expression, thereby upregulating the expression of miR-616's targets SOCS4 (SOCS5). Additionally, in vivo experiments using a xenograft tumor mouse model revealed that TUSC7 can serve as a tumor suppressor through sponging miR-616, and upregulating SOCS4 (SOCS5) in EC. CONCLUSIONS: In this study, a newly identified regulatory mechanism of lncRNA TUSC7/miR-616/ SOCS4 (SOCS5) axis was systematically studied, which may hold promise as a promising target for EC treatment.

Metformin is associated with reduced cell proliferation in human endometrial cancer by inbibiting PI3K/AKT/mTOR signaling.

Metformin recently gained traction as potential anti-endometrial cancer agent for its new applications. However, the underlying mechanisms of the anti-cancer effect of metformin in the endometrial cancer have not yet been fully elucidated. Sixty-five patients diagnosed as endometrial carcinoma were grouped into (n = 33) and non-treatment mixed (n = 32) for analysis. Thirty healthy donors were recruited as controls. We attempt to investigate the effect of metformin on Ki-67, PI3K, p-AKT, p-S6K1, and p-4EBP1 staining in human endometrial cancer by immunohistochemical staining. We found that increased Ki-67 expression in women with endometrial cancer, which were reversed by conventional anti-diabetic doses of metformin in present work. In parallel, the reduced PI3K, p-AKT, p-S6K1, and p-4EBP1 staining induced by metformin appeared to play an important role for the anti-proliferative effects of metformin in endometrial cancer patients. Metformin significantly decreased proliferation in human endometrial cancer may by inhibiting PI3K/AKT/mTOR signaling. Our present results add to the growing body of evidence supporting metformin as a potential anti-cancer agent in endometrial cancer.

[Effects of prenatal restraint stress on neural stem cell proliferation and nestin protein expression in the hippocampus of offspring rats].

To investigate the effect of prenatal stress on neural stem cell proliferation and nestin protein expression in the hippocampus of rat offspring, Sprague-Dawley female rats weighing 240-260 g were included and randomly divided into control and restraint stress groups. Rats in the control group were given no interventions. Rats in the restraint stress group were given restraint stress three times daily for 45 min during gestational days 14-20. One-month-old offspring rats were used for further experiment. The results of Morris water maze showed that the escape latency to reach the platform in offspring of restraint stress group was longer than that in control group (P < 0.05). There was no significant difference of the staying-time in quadrant of the primary platform in spatial probe test. The expressions of nestin- and BrdU-positive cells in the hippocampus of female offspring in restraint stress group were significantly higher than those in the control group (P < 0.05), but there was no significant difference in male offspring (P > 0.05). These results suggest that prenatal restraint stress can increase the number and proliferative capability of the neural stem cells in female offspring. It may be a primary stage of the cascade reaction of the body to the brain damage caused by prenatal restraint stress.

Matrine inhibits the metastatic properties of human cervical cancer cells via downregulating the p38 signaling pathway.

Matrine is a traditional Chinese herbal medicine that shows antitumor efficacy for many types of cancer. The present study evaluated the antitumor efficacy of matrine on cervical cancer and to investigate the underlying mechanisms. We performed MTT assays, and in vitro invasion and migration assays, and P1 L6 found that matrine significantly inhibited cervical cancer cell growth by inducing apoptosis, and suppressed the invasion and migration ability of cervical cancer cells in vitro in a concentration-dependent manner. Mechanistically, we found that matrine decreased the expression and activity of the extracellular matrix factors, matrix metalloproteinases-2 (MMP-2) and MMP-9 via the suppression of p38 signaling pathway. In addition, when cervical cancer cells were grown as xenografts in nude mice, intraperitoneal (i.p.) injection of matrine induced a significant dose-dependent decrease in tumor growth. Taken together, these findings suggest that a potential mechanism by which matrine inhibits the growth and metastasis of cervical cancer through downregulating the p38 signaling pathway.

The homologous recombination protein RAD51 is a promising therapeutic target for cervical carcinoma.

RAD51 is one of the pivotal enzymes for DNA double-strand break (DSB) repair by the homologous recombination (HR) pathway, which implies it as a promising and novel target for cancer therapy. Recent findings have indicated RAD51 protein is overexpressed in a variety of tumors. The high-expression of RAD51 is related to poor prognosis. RAD51 is involved in the repair of DNA damage and the generation of genetic diversity by an evolutionarily conserved mechanism. However, the exact mechanism of Rad51 in the progression of cervical cancer remains unclear. RI-1 is a small molecule that inhibits the central recombination protein RAD51. In this study, we found that RAD51 was highly expressed in invasive squamous cervical cancer (SCC). The administration of RI-1 inhibited cell growth in vitro and reduced growth of tumor xenografts in vivo with cervical cancer cells (HeLa and SiHa). Further investigation suggested that RAD51 protein significantly promoted the cell cycle transition from the G0/G1 to S phase. In addition, the inhibition of RAD51 reduced the level of the cell cycle related protein cyclin D1, but increased the levels of p21 mRNA and protein. As a DNA DSB repair enzyme, the expression of RAD51 in tumor cells possibly affects their sensitivity to anti-cancer agents. Additionally, in experiments using cisplatin and ionizing radiation, RI-1 treated cervical cancer cells, HeLa and SiHa, were sensitized to a greater extent than the untreated control. Thus, HR inhibition of RAD51 may provide yet another mechanism of therapeutic target for the chemosensitization and radiosensitization of cervical cancer with RI-1. Collectively, our data demonstrated for the first time that inhibition of RAD51 suppressed the cervical cancer cell proliferation and the growth of cervical cancer xenografts by attenuating cell cycle transition, which could be a functional link between RAD51 and cyclin D1 and p21.

Maternal PTSD following Exposure to the Wenchuan Earthquake Is Associated with Impaired Mental Development of Children.

The purpose of this study was to explore whether earthquake-related maternal Post-Traumatic Stress Disorder (PTSD) is associated with impaired development of infants. Participants included 86 women who were pregnant during or after the earthquake in Ningqiang county, and their children. Data were collected from February to March of 2012. PTSD questionnaire (PTSD Checklist, Civilian Version (PCL-C)) was used to measure the effect of the earthquake on mothers, and that the scores greater than 50 were used to indicate presence of PTSD. Each child was assessed using the mental Developmental Screening Test (DST) according to age. Among the 86 women, PTSD scores equal to or greater than 50 accounted for 20.93%. Among the 86 children, 25.60% of development quotient (DQ) scores and 19.80% of mental index (MI) scores were less than 85. The correlation coefficient analysis showed that PTSD scores were inversely related to DQ and MI scores. Maternal PTSD following earthquake exposure is associated with relatively lower intellectual development in children age 0-3 years. Further research is needed to assess the persistent effects of this influence on offspring of mothers exposed to earthquake.

Association of a common variant of SYNPO2 gene with increased risk of serous epithelial ovarian cancer.

In China, the majority of ovarian cancer patients (80%-90%) are women who are diagnosed with epithelial ovarian cancer. The SYNPO2 gene has recently been reported to be associated with epithelial ovarian cancer in Europeans. To investigate the association of common variants of SYNPO2 gene with epithelial ovarian cancer in Han Chinese individuals, we designed a case-control study with 719 epithelial ovarian cancer patients and 1568 unrelated healthy controls of Han Chinese descent. A total of 49 tagging single-nucleotide polymorphisms were genotyped; single-single-nucleotide polymorphism association, imputation, and haplotypic association analyses were performed. The single-nucleotide polymorphism rs17329882 was found to be strongly associated with serous epithelial ovarian cancer and with ages ≤49 years, consistent with the pre-menopausal status of analyzed epithelial ovarian cancer cases. Odds ratios and 95% confidence intervals provided evidence of the risk effects of the C allele of the single-nucleotide polymorphism on epithelial ovarian cancer. Imputation analyses also confirmed the results with a similar pattern. Additionally, haplotype analyses indicated that the haplotype block that contained rs17329882 was significantly associated with epithelial ovarian cancer risk, specifically with the serous epithelial ovarian cancer subtype. In conclusion, our results show that SYNPO2 gene plays an important role in the etiology of epithelial ovarian cancer, suggesting that this gene may be a potential genetic modifier for developing epithelial ovarian cancer.

Dominant factors affecting reproductive outcomes of fertility-desiring young women with intrauterine adhesions.

PURPOSE: To identify the dominant factors affecting reproductive outcomes of fertility-desiring young women with intrauterine adhesions (IUA) after hysteroscopic adhesiolysis. METHODS: This prospective observational study included 104 cases of women with IUA. Logistic regression analysis was applied to analyze the value of the location and extent of adhesions, the number of previous uterine cavity surgery, menstrual patterns, and second look hysteroscopy time. RESULTS: In the total 104 patients with IUA, 91 patients (87.5%) had a history of at least one previous uterine cavity surgery. Hypomenorrhoea and amenorrhoea were two most common presenting menstrual abnormalities occurring in 59 (56.7%) and 28 (26.9%) cases, respectively. The clinical pregnancy rate was 60.6% (63/104) and the live birth rate was 39.4% (41/104). Pregnancy rate was higher in patients having mild IUA (81.5%) as compared to moderate (61.3%) or severe type (20.0%) (p < 0.001). The bivariate and binary logistic regression analysis revealed that the location and extent of adhesions were the independent parameters related to the reproductive outcome after hysteroscopic adhesiolysis for fertility-desiring women with IUA (p = 0.011 and p = 0.003, respectively), but not the number of previous uterine cavity surgery, menstrual patterns, and second look hysteroscopy time (p = 0.232, p = 0.239 and p = 0.120, respectively). CONCLUSION: The extent and location of IUA are the dominant factors affecting reproductive outcomes, which are possibly regarded as a potential predictor evaluating prognosis of IUA after hysteroscopic adhesiolysis.

Insulin-Like Growth Factor 1/Mammalian Target of Rapamycin and AMP-Activated Protein Kinase Signaling Involved in the Effects of Metformin in the Human Endometrial Cancer.

BACKGROUND: Metformin is a well-tolerated biguanide drug used for decades to treat type 2 diabetes mellitus. In recent years, long-term administration of metformin has been found to reduce carcinogenic risk for cancers derived from various tissues. However, its cellular and molecular mechanisms of anticancer action in the endometrial cancer (EC) have not yet been fully elucidated. PATIENTS AND METHODS: Sixty patients diagnosed as endometrial carcinoma were grouped into (n = 30) and non-treatment mixed (n = 30) for analysis. Thirty healthy donors are control groups. We attempt to investigate the interaction of metformin, insulin-like growth factor 1 (IGF-1) expression, and phosphorylated mammalian target of rapamycin (p-mTOR) and AMP-activated protein kinase (p-AMPK). RESULTS: We found that high IGF-1 plasma concentrations in women with EC were reversed by conventional antidiabetic doses of metformin in the present work. In parallel, the activation of AMPK and suppression of mTOR seemed to play an important role for the effect of metformin in patients with EC. CONCLUSIONS: This pilot trial presents biological evidence consistent with antiproliferative effects of metformin in women with EC in the clinical setting.

miR-99a and -99b inhibit cervical cancer cell proliferation and invasion by targeting mTOR signaling pathway.

MicroRNAs were demonstrated to play an important role in the regulation of gene expression. Here, we showed that miR-99a and -99b (miR-99a/b) were down-regulated in human cervical cancer patient tissues and were negatively related with lymphatic metastasis. In addition, overexpression of miR-99a/b inhibited cell growth and invasion, whereas suppression of miR-99a/b yielded the reverse phenotype. Dual luciferase report assay revealed that mTOR was identified as a novel target gene of both miR-99a and -99b. Altogether, these results suggested that miR-99a/b directly and negatively regulated mTOR expression in cervical cancer cells, and enforced the importance of miR-99a/b and their targets in the malignant phenotypes of cervical carcinogenesis.

The involvement of ERK/CREB/Bcl-2 in depression-like behavior in prenatally stressed offspring rats.

A number of studies reveal that prenatal stress (PS) may induce an increased vulnerability to depression in offspring. Some evidences indicate that extracellular signal-regulated kinase (ERK)-cyclic AMP responsive element binding protein (CREB) signal system may play an important role in the molecular mechanism of depression. In the present study, we examined the effects of prenatal restraint stress on depression-like behavior in one-month offspring Sprague-Dawley rats and expression of ERK2, CREB, B-cell lymphoma-2 (Bcl-2) mRNA in the hippocampus, prefrontal cortex and striatum to explore the potential role of ERK-CREB pathway in mediating the behavioral effects of PS exposure. Our findings demonstrated that PS increased immobility time in forced swimming test and decreased expression of ERK2, CREB, Bcl-2 mRNA in the hippocampus and prefrontal cortex of juvenile offspring rats except for CREB in hippocampus of male offspring. Changes induced by PS were partly prevented by MK-801, an N-methyl-D-aspartate (NMDA) receptor antagonist. These findings suggested that the ERK-CREB system might be related with the depression-like behavior in juvenile offspring rats subjected to PS, in which NMDA receptors might be involved.