[Using Immunohistochemical Markers and Clinicopathological Factors to Predict the Prognostic Survival of Different Types of Endometrial Cancer Recurrence].

OBJECTIVE: To probe for factors that can be used effectively to predict the prognostic survival of patients with endometrial cancer recurrence. METHODS: The clinicopathological data of 473 patients with stage Ⅰ to Ⅲ endometrial cancer who underwent standard surgical treatment from October 2013 to May 2019 were retrospectively collected, and post-operative recurrence of the patients were followed up. Overall recurrence includes local recurrence and poor prognosis recurrence. The endpoint indicators of this study are the recurrence-free survival (RFS) and overall survival (OS) of patients with overall recurrence, local recurrence, and poor prognosis recurrence (PPR). The Kaplan-Meier survival curve was used to evaluate the OS and RFS of patients. Cox proportional-hazards model was used to identify factors affecting the prognostic survival of patients with endometrial cancer recurrence. RESULTS: Among the 473 patients, 406 did not experience recurrence. A total of 67 patients, accounting for 14.2%, had recurrence. Among them, 27 had local recurrence, accounting for 5.7%, while 40 had poor prognosis recurrence, accounting for 8.5%. The median follow-up time of patients with recurrence was 38 months. The survival curve showed that the RFS and OS of the patients in the recurrence-free group remained unchanged, while the patients in the recurrence group, regardless of whether they had overall recurrence, local recurrence or PPR, experienced a decrease in RFS and OS( P<0.001). The overall 3-year OS rate of patients with recurrence was 44.8%, the median survival time was 29 months, and the median recurrence time was 17 months. The 3-year OS rate of patients in the recurrence-free group was 98.8%, and the median survival time was 40 months; the 3-year OS rate of patients with local recurrence was 59.3%, the median survival time was 27 months, and the median recurrence time was 15 months. The 3-year OS rate of patients with PPR was only 35.0%, the median survival time was 22 months, and the median recurrence time was 10 months. The results of multivariate Cox regression analysis showed that, for overall recurrence patients, FIGO stage Ⅲ (hazard ratio ( HR)=3.432, P=0.005), increased expression of K-i67 ( HR=1.015, P=0.025), and decreased expression of estrogen receptor (ER) ( HR=0.985, P=0.005) are independent factors for the decline in RFS, FIGO stage Ⅲ ( HR=4.918, P=0.005) and the decreased expression of progesterone receptor (PR) ( HR=0.977, P=0.003) are independent factors for the decrease in OS. For patients with local recurrence, special pathological types ( HR=2.545, P=0.049) and increased expression of Ki-67 ( HR=1.024, P=0.033) are independent factors influencing the decrease in RFS, while decreased expression of PR ( HR=0.973, P=0.009) is an independent risk factor for decreased OS. For patients with PPR, FIGO stage Ⅲ ( HR=5.977, P=0.002) and decreased ER expression ( HR=0.984, P=0.023) are independent risk factors for the decline in RFS, while FIGO stage Ⅲ ( HR=10.098, P=0.001) is an independent factor influencing the decline of OS. CONCLUSION: FIGO stage Ⅲ, increased Ki-67 expression, and decreased ER expression can increase patients' risk of postoperative recurrence, and FIGO stage Ⅲ and decreased expression of PR can increase the risk of death in patients with recurrence.

Stomatin-like protein 2 is involved in endometrial stromal cell proliferation and differentiation during decidualization in mice and humans.

The molecular mechanisms underlying endometrial stromal cell proliferation and differentiation (decidualization) are still not fully understood. This study revealed that increased Slp-2 expression is a significant factor modulating endometrial stromal cell proliferation and decidualization in both mice and humans. Our results showed a significant difference in the mRNA and protein levels between the implantation site and inter-implantation site on day 5 and day 6 of pregnancy in mice (all P < 0.05). Strong Slp-2 immunostaining was mainly localized within the decidual zone of mice through the post-implantation period. Mice with artificially induced deciduoma showed significantly higher expression of Slp-2 compared with uninduced controls (P < 0.005). Human stromal cells in the middle and late-secretory phases demonstrated significantly (all P < 0.05) upregulated SLP-2, compared with cells in the proliferative phase and early secretory phases. Further analyses of the SLP-2 gene knocked down revealed a significant (P < 0.005) repression of both the decidualization marker gene's expression (decidual/trophoblast prolactin-related protein in mice, insulin-like growth factor binding protein and prolactin in human) and the cell proliferation in in vitro-induced decidualized primary endometrial stromal cells in mice and humans.

Methylated oligonucleotide (MON)-induced promoter hypermethylation is associated with repression of CDH1 expression and contributes to the migration and invasion of human trophoblast cell lines.

DNA cytosine-5 methylation plays a vital role in regulating the expression of E-cadherin, which is encoded by the CDH1 gene. In this study, we characterised the DNA methylation and expression pattern of CDH1 in an extravillous trophoblast cell line (HTR-8/SVneo) and two trophoblast cell lines -- JEG-3 and JAR. Promoter hypermethylation with reduced E-cadherin expression in HTR-8/SVneo cells and promoter hypomethylation with increased E-cadherin expression in JEG-3 and JAR cells were observed. Demethylation treatment significantly restored E-cadherin expression, contributing to decreases in the motility and invasiveness of HTR-8/SVneo cells. Sense-methylated oligonucleotides (MONs) labelled with Cy5 and complementary to a region of the human CDH1 promoter were designed, with the cytosines in 5'-cytosine-phosphate-guanine-3' (CpG) dinucleotides being replaced by methylated cytosines. Following MON transfection into JEG-3 cells, the level of CDH1 promoter DNA methylation as well as cell motility and invasiveness were increased and gene expression was significantly repressed. Our results indicate that MON-mediated DNA methylation of the CDH1 promoter and subsequent alterations in gene expression may contribute to trophoblast motility and invasion, suggesting a potential method for controlling the biological function of trophoblasts in vitro through epigenetic modification.

nm23 regulates decidualization through the PI3K-Akt-mTOR signaling pathways in mice and humans.

STUDY QUESTION: Does nm23 have functional significance in decidualization in mice and humans? SUMMARY ANSWER: nm23 affects decidualization via the phosphoinositide 3 kinase/mammalian target of rapamycin (PI3K-Akt-mTOR) signaling pathways in mouse endometrial stromal cells (ESCs; mESCs) and human ESCs. WHAT IS KNOWN ALREADY: The function of nm23 in suppressing metastasis has been demonstrated in a variety of cancer types. nm23 also participates in the control of DNA replication and cell proliferation and differentiation. STUDY DESIGN, SIZE AND DURATION: We first analyzed the expression profile of nm23 in mice during early pregnancy (n = 6/group), pseudopregnancy (n = 6/group) and artificial decidualization (n = 6/group) and in humans during the menstrual cycle phases and the first trimester. We then used primary cultured mESCs and a human ESC line, T-HESC, to explore the hormonal regulation of nm23 and the roles of nm23 in in vitro decidualization, and as a possible mediator of downstream PI3K-Akt-mTOR signaling pathways. PARTICIPANTS/MATERIALS, SETTINGS AND METHODS: We evaluated the dynamic expression of nm23 in mice and humans using immunohistochemistry, western blot and real-time quantitative RT-PCR (RT-qPCR). Regulation of nm23 by steroid hormones was investigated in isolated primary mESCs and T-HESCs by western blot. The effect of nm23 knockdown (using siRNA) on ESC proliferation was analyzed by 5-ethynyl-2'-deoxyuridine staining (EdU) and proliferating cell nuclear antigen protein (PCNA) expression. The influence of nm23 expression on the differentiation of ESCs was determined by RT-qPCR using the mouse differentiation markers decidual/trophoblast PRL-related protein (dtprp, also named prl8a2) and prolactin family 3 subfamily c member 1 (prl3c1) and the human differentiation markers insulin-like growth factor binding protein 1 (IGFBP1) and prolactin (PRL). The effects of nm23 siRNA (si-nm23) and the PI3K inhibitor LY294002 on the downstream effects of nm23 on the PI3K-Akt-mTOR signaling pathway were estimated by western blot. MAIN RESULTS AND THE ROLE OF CHANCE: NM23-M1 was specifically expressed in the decidual zone during early pregnancy and in artificially induced deciduoma, and NM23-H1 was strongly expressed in human first trimester decidua. The expression of nm23 was upregulated by oestradiol and progesterone (P < 0.05 versus control) in vitro in mESCs and T-HESC, and this was inhibited by their respective receptor antagonists, ICI 182,780 and RU486. Mouse and human nm23 knockdown decreased ESC proliferation and differentiation (P < 0.05 versus control). The PI3K-Akt-mTOR signaling pathways were downstream mediators of nm23 in mESCs and T-HESCs decidualization. LIMITATIONS AND REASONS FOR CAUTION: Whether the nm23 regulates decidualization via the activation of AMPK, RAS, PKA, STAT3 or other signaling molecules remains to be determined. The role of nm23 in decidualization was tested in vitro only. WIDER IMPLICATIONS OF THE FINDINGS: Results demonstrate that nm23 plays a vital role in decidualization in mice and humans and that nm23 gene expression is hormonally regulated. The downregulation of nm23 in decidua during the first trimester may be associated with infertility in women. STUDY FUNDING/COMPETING INTERESTS: This study was supported by the National Natural Science Foundation of China (grant nos. 81370731, 31571551 and 31571190), the Science and Technology Project of Chongqing Education Committee (KJ130309), open funding by the Chongqing Institute for Family Planning (1201) and the Excellent Young Scholars of Chongqing Medical University (CQYQ201302). The authors have no conflicts of interest to declare.

Clinicopathological significance of steroidogenic factor-1 expression in ovarian cancer versus ovarian sex cord stromal tumor.

The significance of steroidogenic factor-1 (SF-1) in human ovarian tumor has not been fully investigated. The purposes of this study are to provide a meta-analysis for SF-1 and to determine whether SF-1 is associated with ovarian tumor progression and clinicopathological characteristics. A detailed literature search was made for related research publications written in English. Methodological quality of the studies was also evaluated. The data were extracted and assessed by two reviewers independently. Analysis of pooled data was performed, and odds ratio (OR) and corresponding confidence intervals (CIs) were calculated and summarized respectively. Final analysis from seven eligible studies was performed. Aberrant SF-1 expression was significantly lower in ovarian cancer compared to that of normal ovarian tissue (OR = 0.02, 95% CI = 0.00-0.16, p = 0.0002). However, SF-1 protein expression was not significantly different between benign and malignant ovarian tumors (p = 0.35). Interestingly, aberrant SF-1 expression was significantly higher in ovarian sex cord stromal tumors than that of ovarian cancer (OR = 0.00, 95% CI = 0.00-0.01, p < 0.00001). The results of this meta-analysis suggest that SF-1 may play an important role in ovarian cancer initiation and progression. Moreover, SF-1 expression may serve as a marker in the differential diagnosis between ovarian sex cord stromal tumors and ovarian cancer.

Aberrant promoter hypermethylation of p16 gene in endometrial carcinoma.

Previous studies demonstrated that the loss of function of the p16INK4A gene is mainly caused by the hypermethylation of p16 gene promoter; however, whether or not it is associated with the incidence of endometrial carcinoma (EC) remains unclear. In the current study, we conducted a meta-analysis to investigate the effects of p16 gene promoter hypermethylation on the incidence of EC. Detailed research publications were searched from Embase, PubMed, and ISI Web of Knowledge for composition in English or Chinese. The pooled data were collected and analyzed by Review Manager 5.2. Odds ratios (ORs) were calculated and summarized respectively. Six eligible studies, including 261 patients were selected and analyzed. The pooled OR was 0.42, test for overall effect, Z = 10.19, P < 0.0001, indicating that p16 gene promoter hypermethylation was significantly correlated with the EC patients. The results of our study strongly suggest that p16 gene promoter hypermethylation is correlated with an increased risk of EC. P16 gene promoter hypermethylation plays a critical role in endometrial carcinogenesis.