Long noncoding RNA H19: functions and mechanisms in regulating programmed cell death in cancer.

Long noncoding RNAs (lncRNAs) are a group of noncoding RNAs with transcript lengths of >200 nucleotides. Mounting evidence suggests that lncRNAs are closely associated with tumorigenesis. LncRNA H19 (H19) was the first lncRNA to function as an oncogene in many malignant tumors. Apart from the established role of H19 in promoting cell growth, proliferation, invasion, migration, epithelial-mesenchymal transition (EMT), and metastasis, it has been recently discovered that H19 also inhibits programmed cell death (PCD) of cancer cells. In this review, we summarize the mechanisms by which H19 regulates PCD in cancer cells through various signaling pathways, molecular mechanisms, and epigenetic modifications. H19 regulates PCD through the Wnt/ß-catenin pathway and the PI3K-Akt-mTOR pathway. It also acts as a competitive endogenous RNA (ceRNA) in PCD regulation. The interaction between H19 and RNA-binding proteins (RBP) regulates apoptosis in cancer. Moreover, epigenetic modifications, including DNA and RNA methylation and histone modifications, are also involved in H19-associated PCD regulation. In conclusion, we summarize the role of H19 signaling via PCD in cancer chemoresistance, highlighting the promising research significance of H19 as a therapeutic target. We hope that our study will contribute to a broader understanding of H19 in cancer development and treatment.

Single-cell transcriptome analysis reveals endometrial immune microenvironment in minimal/mild endometriosis.

Endometriosis is a common inflammatory disorder in women of reproductive age due to an abnormal endometrial immune environment and is associated with infertility. This study aimed to systematically understand the endometrial leukocyte types, inflammatory environment, and impaired receptivity at single-cell resolution. We profiled single-cell RNA transcriptomes of 138 057 endometrial cells from endometriosis patients (n = 6) and control (n = 7), respectively, using 10x Genomics platform. We found that one cluster of epithelial cells that expressed PAEP and CXCL14 was mostly from the control during the window of implantation (WOI). This epithelial cell type is absent in the eutopic endometrium during the secretory phase. The proportion of endometrial immune cells decreased in the secretory phase in the control group, whereas the cycle variation of total immune cells, NK cells, and T cells was absent in endometriosis. Endometrial immune cells secreted more IL-10 in the secretory phase than in the proliferative phase in the control group; the opposite trend was observed in endometriosis. Proinflammatory cytokines levels in the endometrial immune cells were higher in endometriosis than in the control group. Trajectory analysis revealed that the secretory phase epithelial cells decreased in endometriosis. Ligand-receptor analysis revealed that 11 ligand-receptor pairs were upregulated between endometrial immune and epithelial cells during WOI. These results provide new insights into the endometrial immune microenvironment and impaired endometrial receptivity in infertile women with minimal/mild endometriosis.

Existence of chronic endometritis and its influence on pregnancy outcomes in infertile women with minimal/mild endometriosis.

OBJECTIVE: This study aimed to evaluate the prevalence of chronic endometritis (CE) in women with minimal/mild endometriosis and to analyze whether CE affects their pregnancy outcomes. METHODS: This retrospective study included 201 infertile women who were diagnosed with minimal/mild endometriosis after undergoing hysteroscopy combined with laparoscopy from January 2016 to December 2018. Immunohistochemistry was used to detect CD138 and CD38, which are specific markers of plasma cells in the endometrial stroma to diagnose CE. Subsequently, we investigated the prevalence of CE and the effects of CE on spontaneous cumulative pregnancy rate, live birth rate, and miscarriage rate within 24 months after surgery. RESULTS: The prevalence of CE in infertile women with minimal/mild endometriosis was 24.38%. Patients diagnosed with CE showed a significantly lower cumulative pregnancy rate and live birth rate compared with women without CE (46.51% vs. 71.13% [P = 0.004]; 44.19% vs. 63.38% [P = 0.025]). However, the rate of miscarriage in women with CE was also lower than in women without CE (0 vs. 7.04%, P = 0.074). CONCLUSION: Since CE had an adverse effect on cumulative pregnancy rate and live birth rate in infertile women with minimal/mild endometriosis, we suggested that diagnosis and treatment of CE may improve their pregnancy outcomes.

miR-297 inhibits expression of progesterone receptor and decidualization in eutopic endometria of endometriosis.

AIM: Progesterone resistance is an epigenetic factor that reduces endometrial receptivity and causes implantation failure in women with endometriosis. In addition, dysregulated miRNAs contribute to the underlying pathogenic mechanisms of endometriosis. This study aimed to determine the effect of miR-297 on the progesterone receptor (PR) expression and on insufficient decidualization of endometrial stromal cells (ESCs) within the eutopic endometria of infertile women with minimal or mild endometriosis. METHODS: ESCs were isolated from infertile endometriosis and normal patients and were transfected with miR-297 mimic or miR-297 inhibitor or respective control. qRT-PCR and western blot were conducted to quantify the expression of miR-297 and PR. The effect of miR-297 on ESCs decidualization was investigated by induced decidualization in vitro. RESULTS: We observed an increase in miR-297 expression and a decrease in the expression of PR in the ESCs from endometriosis patients. Moreover, the expression of PR, most notably PRB, was found to be downregulated following transfection with miR-297 mimic and upregulated following treatment with miR-297 inhibitor. In addition, overexpressed miR-297 inhibited the decidualization of ESCs in vitro. We further determined that miR-297 exerts direct regulatory effects on PR expression. CONCLUSIONS: We demonstrated that miR-297 interferes with fertility by repressing the expression of PR and preventing efficient decidualization in eutopic endometria. Further, miR-297 directly contributes to progesterone resistance in minimal or mild cases of endometriosis. Thus, regulation of miR-297 may prove to be a promising therapeutic strategy for endometriosis.

Increased Expression of YAP Inhibited the Autophagy Level by Upregulating mTOR Signal in the Eutopic ESCs of Endometriosis.

We first reported that the Hippo-YAP signaling pathway plays a critical role in the pathogenesis of endometriosis (EMS). Autophagy is also related to the invasion ability of endometrial cells and is involved in the pathogenesis of EMS through multi-levels. However, the precise regulatory mechanism of YAP on autophagy in the eutopic endometrial stromal cells (ESCs) is still unclear. Primary eutopic ESCs of EMS patients (n = 12) and control patients without EMS (n = 9) were isolated and cultured to investigate the expressions of YAP and mTOR, the role of YAP in autophagy, and the effect of the YAP-autophagy signal on the decidualization of the eutopic ESCs. Endometriosis-related sequencing data (GSE51981) in the GEO database were used to find the genes significantly correlated with YAP. We found 155 genes with significant differences in the interaction with YAP in EMS from the dataset, and the autophagy pathway was significantly enriched. Following on from our previous studies of YAP knockdown, overexpression of YAP resulted in an increased expression of mTOR and decreased ratio of LC3-II/LC3-I and autophagy markers, in the eutopic ESCs; transmission electron microscope observation also showed fewer autophagosomes compared with the control cells. Furthermore, ESCs of the Rapamycin-treated group showed significant decidual-like changes with significantly increased decidual prolactin level at 72 h after in vitro decidualization. These results demonstrate that the increased YAP inhibited the level of autophagy by upregulating the mTOR signal in the eutopic ESCs of endometriosis. The YAP-autophagy signal plays an important role in the pathogenesis of endometriosis-associated infertility.

Comparative Proteomic Analysis Reveals Metformin Improves the Expression of Biomarkers of Endometrial Receptivity in Infertile Women with Minimal/Mild Endometriosis.

The prevalence of endometriosis is approximately 10% in women of reproductive age, and 30-50% of women with endometriosis are infertile. Metformin has been reported to inhibit the growth of ectopic lesions in endometriosis. However, its effect on the eutopic endometrium of endometriosis is unknown. This study aimed to identify whether metformin affects endometrial receptivity in infertile women with minimal/mild endometriosis. We enrolled 10 infertile women who were diagnosed with minimal/mild endometriosis through laparoscopy. Paired endometrial tissues of the secretory phase from participants were collected during surgery and after 2 months of metformin treatment (n = 5) or no medical treatment (n = 5). Protein expression profiles of the paired endometrium were detected by proteomics and compared using the self-control method (2 months later vs. in surgery). Proteomics data revealed six proteins associated with endometrial receptivity among the significantly upregulated proteins after metformin treatment (fold change > 1.5, P < 0.05). Insulin-like growth factor binding protein 7 (IGFBP-7) showed the most robust increase in these six endometrial receptivity-related proteins (fold change: 8.668, P < 0.05), while there was no significant change in the controls (fold change: 1.906, P > 0.05). The upregulation of IGFBP-7 has been validated through target proteomics, immunohistochemistry, and further demonstrated in endometriosis mouse models induced by autotransplantation. This study revealed that metformin upregulated the expression of IGFBP-7 in the endometrium of human and mouse models of endometriosis. Metformin potentially affects endometrial receptivity of minimal/mild endometriosis by improving the expression of the endometrial receptivity marker IGFBP-7.

Androgen levels in the fetal cord blood of children born to women with polycystic ovary syndrome: a meta-analysis.

INTRODUCTION: Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders in reproductive-aged women. It is reported that intrauterine exposure to hyperandrogenism may induce the development of PCOS and associated complications in later life. To analyze the intrauterine androgen levels in infants born to PCOS mothers, we evaluated the androgen levels in fetal cord blood through a meta-analysis of observational studies. MATERIAL AND METHODS: The following online databases were systematically searched: PubMed, EMBASE, Cochrane library databases and Web of Science up to December 2019. Human studies compared cord blood androgen levels, including testosterone (T) and androstenedione (ADION), in fetal cord blood of mothers with and without PCOS. Statistical analysis was performed in Review Manager, Version 5.3, with the inverse variance method based on a random-effects model. RESULTS: A total of 7 articles were scrutinized and a total of 570 samples including 268 female and 222 male infants were qualified for review. In the mass spectrograph (MS) subgroup, PCOS mothers showed no signs of increased T concentration in umbilical cord blood at birth (4 studies; hazard ratio [HR] = - 0.05; 95% confidence interval [CI] = [- 0.33,0.24]; I2 = 7%; P = 0.75; fixed-effects model). ADION level tends to be lower in daughters' cord blood of PCOS mothers (3 studies; HR = -0.59; 95%CI = [- 1.00, - 0.19]; I2 = 0%; P = 0.004; fixed-effects model). CONCLUSIONS: Fetal cord blood T level is not related to PCOS, while ADION levels tend to be lower in the cord blood of daughters born to mothers with PCOS.

MicroRNA22-5p targets ten-eleven translocation and regulates estrogen receptor 2 expression in infertile women with minimal/mild endometriosis during implantation window.

Based on microRNA (miR) microarray analysis, we previously found that miR22-5p expression is decreased in the mid-luteal endometrium of women with minimal/mild endometriosis. Bioinformatics analysis predicted that miR22-5p targets ten-eleven translocation (TET2) 3'-untranslated region. This study aimed to determine the regulation and roles of miR22-5p in the pathogenesis of minimal/mild endometriosis-associated infertility. MiR22-5p and TET2 expression in the mid-luteal endometrium from women with or without minimal/mild endometriosis was analyzed. After transfection with miR22-5p mimics or inhibitor, TET2 expression was analyzed by quantitative reverse transcription (RT-q) PCR, western blotting and immunohistochemistry. 5-Hydroxymethylcytosine was determined by immunofluorescence and dot blotting. Expression and promoter methylation of estrogen receptor 2 (ESR2) was measured by RT-qPCR and western blotting, and by bisulfite sequencing, respectively. We first established that miR22-5p expression decreased and TET2 expression increased in minimal/mild endometriosis during implantation window. TET2 was found to be a direct target of miR22-5p. MiR22-5p regulated the expression of ESR2, but did not directly affect methylation of its promoter region (-197/+359). Our results suggest that an imbalance in miR22-5p expression in the mid-luteal endometrium may be involved in minimal/mild endometriosis-associated infertility.

Clinicopathological and Prognostic Characteristics of CD276 (B7-H3) Expression in Adrenocortical Carcinoma.

BACKGROUND: Adrenocortical carcinoma (ACC) is a rare malignant endocrine tumor with a high tumor recurrence rate and poor postoperative survival. Recent studies suggest that CD276- (B7-H3) targeted therapy represents a promising therapeutic option for solid tumors. However, little is known about the expression status of CD276 or its association with progression and prognosis of ACC. METHODS: Clinical data were retrospectively analyzed from patients who underwent resection of ACC at our institution (n = 48). Archived, formalin-fixed, and paraffin-embedded samples were collected for immunohistochemical analysis, and the correlation between CD276 expression and clinicopathological parameters was evaluated. Kaplan-Meier and univariate/multivariate Cox regression methods were implemented to identify any prognostic effects. Data from The Cancer Genome Atlas (TCGA) ACC cohort (n = 48). Archived, formalin-fixed, and paraffin-embedded samples were collected for immunohistochemical analysis, and the correlation between CD276 expression and clinicopathological parameters was evaluated. Kaplan-Meier and univariate/multivariate Cox regression methods were implemented to identify any prognostic effects. Data from The Cancer Genome Atlas (TCGA) ACC cohort (. RESULTS: Positive expression of CD276 was detected on the cell membrane and in the cytoplasm of cancer cells or tumor-associated vascular cells in 91.67% (44/48) of ACCs. Vascular expression of CD276 was associated with local aggression (higher T stage, P = 0.029) and advanced ENSAT stage (P = 0.029) and advanced ENSAT stage (P = 0.029) and advanced ENSAT stage (P = 0.029) and advanced ENSAT stage (P = 0.029) and advanced ENSAT stage (P = 0.029) and advanced ENSAT stage (. CONCLUSION: These findings highlight the immune checkpoint factor CD276 as an independent prognostic factor and a potential therapeutic target in ACC.

Increased expression of YAP is associated with decreased cell autophagy in the eutopic endometrial stromal cells of endometriosis.

Basic research has shown that signal pathways play significant roles in the development and progression of endometriosis (EMS). We first reported that the Hippo-YAP (Yes-associated protein) pathway promotes cell proliferation and anti-apoptosis in endometrial stromal cells (ESCs) of EMS. Cell autophagy has been found to be involved in the endometrial regulation and the pathophysiology of EMS. We speculated that there may be an elaborate dialogue between Hippo-YAP and autophagy pathway in EMS. To explore this, we performed molecular biology experiments to investigate the expressions of YAP pathway and cell autophagy markers (mTOR, LC-3) in ESCs of women with or without EMS and detected the protein levels of autophagy markers after verteporfin and rapamycin treatments and the transfection with YAP-knockdown vector in the eutopic ESCs, respectively. We found that the mRNAs of YAP and mTOR were increased in the eutopic ESCs compared with controls, but no statistically difference, while the protein levels were significantly increased in the eutopic ESCs. Conversely, the ratio of the autophagy marker protein LC3-II/LC3-I was significantly decreased in the eutopic ESCs compared with controls. Moreover, verteporfin treatment interfered with the YAP function, but it had no effect on mTOR expression and cell autophagy level. Rapamycin treatment and YAP knockdown in the eutopic ESCs both inhibited the expression of YAP and increased the ratio of LC3-II/LC3-I significantly. These results demonstrate that the decreased cell autophagy level is associated with the increased expression of YAP and YAP may participate in the mTOR-autophagy pathway in the eutopic ESCs of endometriosis.

Body fat indices as effective predictors of insulin resistance in obese/non-obese polycystic ovary syndrome women in the Southwest of China.

PURPOSE: Insulin resistance (IR) is a common feature of polycystic ovary syndrome (PCOS). Body fat indices can be predictive markers of IR. This study is aimed to predict IR in Chinese women with PCOS of different body types based on body fat indices. METHODS: A total of 723 women diagnosed with PCOS according to Rotterdam criteria were recruited in this study and were further divided into two groups based on their BMI. All participants underwent physical examinations and ultrasound; and blood was collected from them on the days 3-5 of the menstrual cycle. Their BMI, waist-to-hip ratio (WHR), waist-to-height ratio (WHtR), lipid accumulation product (LAP) index, visceral adiposity index (VAI), and the homeostasis model assessment index of insulin resistance (HOMA-IR) were calculated. The correlations between body fat indices and HOMA-IR and receiver operating characteristic (ROC) curves were evaluated. RESULTS: In normal weight group (BMI < 24, n = 333), VAI (best cut-off value: 1.681, area under curve (AUC) = 0.754, P < 0.01) and LAP index (best cut-off value: 18.53, AUC = 0.734, P < 0.001) were the reliable indicators of IR based on HOMA-IR ≥ 2.77, while in overweight/obese group (BMI ≥ 24, n = 390), the BMI, WC, WHtR and LAP index had a significant correlation with HOMA-IR. The representative markers to assess IR were BMI (best cut-off value: 26.43, AUC = 0.644, P = 0.001) and WHtR (best cut-off value: 0.544, AUC = 0.604, P = 0.021). CONCLUSIONS: Body fat indices are predictive markers of IR in Chinese PCOS women, especially in those with normal weight.

Anthropometric indices to predict insulin resistance in women with polycystic ovary syndrome in China.

RESEARCH QUESTION: Which anthropometric index (waist-to-height ratio, waist circumference, waist-to-hip ratio and body mass index) is the best in predicting insulin resistance among Chinese women with polycystic ovary syndrome? DESIGN: A total of 1124 patients with polycystic ovary syndrome at the Reproductive Endocrinology Division of West China Second University Hospital of Sichuan University were enrolled in this study. Identification of insulin resistance was based on homeostasis model assessment of insulin resistance scores 2.77 or over. Receiver operator characteristic analysis was carried out using the four anthropometric indices as the continuous variables and insulin resistance as the categorical variable to obtain the areas under the curve. RESULT: The area under the curve for the waist-to-height ratio (0.748 ± 0.019) was greater than those for waist circumference (0.739 ± 0.019), body mass index (0.738 ± 0.017), and waist-to-hip ratio (0.659 ± 0.020) in the prediction of insulin resistance. The waist-to-height ratio also had the highest Youden indices compared with those of waist circumference, body mass index, and waist-to-hip ratio; the waist-to-height ratio cut-off was 0.49. CONCLUSION: The waist-to-height ratio with a cut-off of 0.49 was the most accurate anthropometric indicator for predicting insulin resistance among Chinese women with polycystic ovary syndrome.

miR-194-3p Represses the Progesterone Receptor and Decidualization in Eutopic Endometrium From Women With Endometriosis.

Progesterone resistance in the eutopic endometrium (EuE) is suggested to be a critical factor for decreased endometrial receptivity and implantation failure in reproductive-aged women with endometriosis. Altered expression of miRNAs has been reported to play an important role in the pathophysiology of endometriosis-associated infertility. However, the underlying mechanisms of aberrant progesterone receptor (PR) and deficient decidualization regulated by miRNAs in endometriosis have not been thoroughly elucidated. The goal of this study was to explore the regulation and roles of miR-194-3p in aberrant PR expression and impaired decidualization in endometrial stromal cells (ESCs) from the EuE of women with mild or minimal endometriosis. Using a series of studies, we observed decreased PR mRNA expression and an increasing PR-A/PR-B mRNA ratio trend in the midsecretory phase of the EuE of women with minimal or mild endometriosis (n = 19) compared with controls (n = 14); the increased expression of miR-194-3p in the endometriosis group was consistent with previous microarray analysis. We also found that PR protein levels were inhibited by the transfection of ESCs with an miR-194-3p mimic and upregulated by miR-194-3p inhibition. As predicted by the bioinformatic analysis, the 3'-untranslated region luciferase assay indicated the direct regulation of PR expression by miR-194-3p. Furthermore, miR-194-3p overexpression inhibited the in vitro decidualization of ESCs via both cellular morphological changes and prolactin levels. Therefore, our study demonstrated that miR-194-3p contributes to progesterone resistance in endometriosis, which hinders fertility by repressing the levels of PR and decidualization in the EuE. Thus, miR-194-3p regulation is a future therapeutic strategy for endometriosis.