HOTAIR/miR-1277-5p/FBN2 signaling axis is involved in recurrent spontaneous abortion by regulating the growth, migration, and invasion of HTR-8/SVneo cells.

OBJECTIVE: This study aimed to explore the specific pathways by which HOX transcript antisense intergenic RNA (HOTAIR) contributes to the pathogenesis of unexplained recurrent spontaneous abortion (URSA). METHODS: Real-time quantitative PCR (RT-qPCR) was employed to assess the differential expression levels of HOTAIR in chorionic villi tissues from URSA patients and women with voluntarily terminated pregnancies. HTR-8/SVneo served as a cellular model. Knockdown and overexpression of HOTAIR in the cells were achieved through siRNA transfection and pcDNA3.1 transfection, respectively. Cell viability, migration, and invasion were evaluated using cell counting kit-8 (CCK-8), scratch, and Transwell assays, respectively. The interaction among the HOTAIR/miR-1277-5p/fibrillin 2 (FBN2) axis was predicted through bioinformatics analysis and confirmed through in vitro experiments. Furthermore, the regulatory effects of the HOTAIR/miR-1277-5p/FBN2 signaling axis on cellular behaviors were validated in HTR-8/SVneo cells. RESULTS: We found that HOTAIR was downregulated in chorionic villi tissues from URSA patients. Overexpression of HOTAIR significantly enhanced the viability, migration, and invasion of HTR-8/SVneo cells, while knockdown of HOTAIR had the opposite effects. We further confirmed the regulatory effect of the HOTAIR/miR-1277-5p/FBN2 signaling axis in URSA. Specifically, HOTAIR and FBN2 were found to reduce the risk of URSA by enhancing cell viability, migration, and invasion, whereas miR-1277-5p exerted the opposite effects. CONCLUSION: HOTAIR promotes URSA development by targeting inhibition of miR-1277-5p/FBN2 axis.

The risk of endometriosis by early menarche is recently increased: a meta-analysis of literature published from 2000 to 2020.

PURPOSE: The relationship between menarche age and endometriosis has been studied extensively; however, the results were inconsistent due to differences in study dates, populations, and methodology. The goal of this meta-analysis was to see how different research populations, dates, and types affected the estimated risk of endometriosis in early menarche. METHODS: The terms "endometriosis", "early menarche", and other pertinent terms were searched in PubMed, Medline, and Embase. This meta-analysis comprised 16 papers published between 2000 and 2020, with a start year ranging from 1989 to 2017. Random effects were used to examine endometriosis risks in these articles. Study start years, countries, types (case-control and cohort studies), and quality (Newcastle-Ottawa scale, NOS) were all taken into account in further stratified analysis and meta-regression analyses. RESULTS: Early menarche (< 12 years) was associated with a significant pooled risk of endometriosis with high heterogeneity (OR = 1.34, 95% CI 1.16-1.54, I2 = 72.0%). Stratified analysis showed that this risk was increased in studies started after 2000 (OR = 1.62, 95% CI 0.96-1.35, I2 = 74.4%), compared to those before 2000 (OR = 1.13, 95% CI 1.16-1.54, I2 = 40.7%); additionally, this risk was higher in low-income countries (OR = 2.11, 95% CI 1.55-2.87, I2 = 0%) than in other countries (OR = 1.25, 95% CI 1.09-1.43, I2 = 70.6%). These results were further confirmed by meta-regression analysis (both P values < 0.1). No significant differences were found between different study types and NOS scores. CONCLUSION: Our results suggested an increasing risk of endometriosis with early menarche, which was more noticeable in low-income countries. Large-scale studies are warranted.

cfDNA Methylation Profiles and T-Cell Differentiation in Women with Endometrial Polyps.

DNA methylation is a part of the regulatory mechanisms of gene expression, including chromatin remodeling and the activity of microRNAs, which are involved in the regulation of T-cell differentiation and function. However, the role of cfDNA methylation in T-cell differentiation is entirely unknown. In patients with endometrial polyps (EPs), we have found an imbalance of T-cell differentiation and an aberrant cfDNA methylation profile, respectively. In this study, we investigated the relationship between cfDNA methylation profiles and T-cell differentiation in 14 people with EPs and 27 healthy controls. We found that several differentially methylated genes (DMGs) were associated with T-cell differentiation in people with EPs (ITGA2-Naïve CD4, r = -0.560, p = 0.037; CST9-EMRA CD4, r = -0.626, p = 0.017; and ZIM2-CM CD8, r = 0.576, p = 0.031), but not in healthy controls (all p > 0.05). When we combined the patients' characteristics, we found a significant association between ITGA2 methylation and polyp diameter (r = 0.562, p = 0.036), but this effect was lost when adjusting the level of Naïve CD4 T-cells (r = 0.038, p = 0.903). Moreover, the circulating sex hormone levels were associated with T-cell differentiation (estradiol-Naïve CD4, r = -0.589, p = 0.027), and the cfDNA methylation profile (testosterone-ZIM2, r = -0.656, p = 0.011). In conclusion, this study has established a link between cfDNA methylation profiles and T-cell differentiation among people with EPs, which may contribute to the etiology of EPs. Further functional studies are warranted.

LIN28B Polymorphisms Confer a Higher Postoperative Recurrence Risk in Reproductive-Age Women with Endometrial Polyps.

The RNA-binding protein LIN28B is an important factor for cell proliferation. Because LIN28B polymorphisms have been shown to be relative with the recurrence of some hyperplastic diseases, we hypothesized that genetic variants of LIN28B gene were associated with postoperative recurrence risk in reproductive-age women with endometrial polyps (EP). In a hospital-based cohort of 351 reproductive female patients underwent hysteroscopic polypectomies between May 2018 and Jan 2020, we genotyped two common polymorphisms in LIN28B gene (rs369065 C > T and rs314280 A > G) and analyzed their associations with the risk of postoperative recurrence in multiple Cox regression model. When followed up to Jun 2021, carries of rs369065 TT genotype had an increased risk of polyp recurrence (adjusting hazard ratio [HR] = 1.883, 95% confidence interval [CI] = 1.033 - 3.434) and had a shorter time to recurrence (median time 352 vs. 342 days, log-rank P < 0.01), compared to the CC/CT genotype. Further stratification analysis showed that the increased risk of rs369065 TT genotype was more evident in patients who were older than 33 years (adjusted HR = 2.597, 95%CI = 1.037 - 6.505), had a single polyp (adjusted HR = 2.545, 95%CI = 1.059 - 6.113), and had smaller polyps (<1.2 cm, adjusted HR = 2.708, 95%CI = 1.042 - 7.043). However, no significant association between rs314280 A > G polymorphism and the risk of polyp recurrence was found. Our study suggests that rs369065 TT genotype of LIN28B gene is associated with an increased postoperative recurrence risk in EP patients, especially in those with fewer and smaller polyps. These findings implicate a precise choice of clinical counseling and decision making. Larger studies in different ethnic populations are warranted.

Circulating PD1+Vδ1+γδ T Cell Predicts Fertility in Endometrial Polyp Patients of Reproductive-Age.

Clinically, immune cell function is correlated with pathogenesis of endometrial polyp (EP) and infertility of women of reproductive-age. However, the underlying immune cell hallmark in EP patients remains unclear. Here, we focused on analyzing circulating immune cells, and attempted to reveal the correlation between peripheral immune cell functional phenotypes and fertility in EP patients. Through comparison of circulating CD4+/CD8+ T cells, NK cells, and γδ T cells between 64 EP patients and 68 healthy females, we found that γδ T cells, but not CD4+/CD8+ T cells and NK cells, were immunologically correlated with conception rate and conception interval time. Specifically, total γδ T cells and the Vδ1+PD1+ γδ T subpopulation decreased whereas the Vδ1/Vδ2 ratio increased in EP patients compared to healthy controls. Moreover, the patients with the higher Vδ1/Vδ2 ratio (median value equals 1.04) had a poorer fertility and longer interval time of conception (210 days versus 158 days for control). Meanwhile, higher Vδ1+PD1+ γδ T cell proportion (median equals 15.7) was positively correlative with both higher conception rate and shortened median conception interval time (130 days for Vδ1+PD1high group versus 194 days for Vδ1+PD1low group). Notably, in healthy controls, both Vδ1/Vδ2 ratio and Vδ1+PD1+ γδ T cell proportion correlated with pregnancy rate oppositely, comparing to EP patients. Together, our results suggested that imbalanced γδ T cell population occurred in EP patients, and that Vδ1/Vδ2 ratio and PD-1 expression of Vδ1+ γδ T cells could be potentially developed into valuable predictors for fertility in EP patients.

Circulating CD56+ NKG2D+ NK cells and postoperative fertility in ovarian endometrioma.

The current biomarkers for postoperative fertility assessment caused by ovarian endometrioma (OE) are insufficient. The present study hypothesized that the peripheral lymphocyte subpopulation can be used as a candidate biomarker of postoperative infertility in OE. The association of the number of circulating CD4/CD8 T, NK, and γδ T cells with postoperative fertility was assessed in 33 OE patients aged 20 ~ 40 years between June 2018 and January 2019. Concomitantly, 68 healthy female subjects were recruited. The changes in the baseline immune characteristics between the two groups were compared. The data demonstrated significant differences in the ratio of CD4/CD8 T cells and the number of CD56+ NKG2D+ NK cells and γδ T cells between OE patients and control subjects. The patients were followed-up till December 2019 and the number of CD56+ NKG2D+ NK cells in the cases was a significant predictor for postoperative fertility as determined by different COX regression models (crude HR = 0.220, 95% CI = 0.059-0.822; adjusted HR = 0.127, 95% CI = 0.024-0.675). A significant delay to successful pregnancy was noted in OE patients (median time, 173 vs. 99 days, log-rank P = 0.013). The present findings suggested that CD56+ NKG2D+ NK cells are a candidate biomarker of postoperative fertility in OE patients. Larger population studies are warranted.

Polymorphisms in XPC Gene and Risk of Uterine Leiomyoma in Reproductive Women.

XPC gene belongs to DNA repair pathway, which is involved in the development of uterine leiomyoma. However, its relationships with leiomyoma risk were never reported. We here hypothesized that XPC gene was associated with the risk of uterine leiomyoma. In this case-control study with a total of 391 leiomyoma cases and 493 tumor-free controls in a reproductive women population in South China, two missense polymorphisms rs2228001 A > C (Lys939Gln) and rs2228000 C > T (Ala499Val) were genotyped by quantitative polymerase chain reaction (qPCR). Then, the associations between these two polymorphisms and leiomyoma risk were investigated. It was revealed that the rs2228000 CT/TT variant genotypes had a decreased leiomyoma risk (adjusted odds ratio = 0.73, 95% confidence interval = 0.54-0.94) compared with rs2228000 CC genotype. Further stratified analysis also revealed that the protective effect of rs2228000 CT/TT on the risk of uterine leiomyoma was more evident among subjects who were younger than 35 years old compared with those with larger tumors (diameter of tumor >5 cm), and those with fewer number of myomas (only one). However, no significant association was observed for leiomyoma risk for rs2228001 A > C. This study indicated that genetic variations in XPC gene are associated with leiomyoma susceptibility in a reproductive women population. It warrants further confirmation in larger prospective studies with different populations.

Characteristics of Peripheral Immune Function in Reproductive Females with Uterine Leiomyoma.

Inflammation and immunity are thought as risk factors for uterine leiomyoma; however, detailed reports on this topic are scarce. The present study aimed to analyze the characteristics of immune function and clinical significance of circulating CD4/CD8 T, NK, and γδ T cells in reproductive females with uterine leiomyoma. We analyzed the above-mentioned cells in 30 reproductive females with uterine leiomyoma and 68 healthy females using flow cytometry. After that, the correlation between function of immune cells and clinical phenotypes was analyzed. Compared with healthy controls, central memory (CM) CD4/CD8 T cells as well as Treg and Tfh cells were notably increased in leiomyoma patients; however, NK and γδ T cells were decreased in patients. Moreover, such alterations of these cells in patients with leiomyoma were associated with shorter menstrual cycles, longer menstrual period, anemia, pelvic lesions, more and larger myomas, and higher levels of CA125. Additionally, the increased Tfh1/Tfh2 ratio and Tfh17 were significantly associated with longer menstrual period, more myomas, and higher CA125 levels independent of age in patients with uterine leiomyoma. In conclusion, hallmarks of peripheral immune function are remarkably correlated with clinical phenotypes in reproductive females with uterine leiomyoma. This preliminary work may provide proof-of-concept for evaluating efficacy of treatment and prognosis of reproductive females with uterine leiomyoma with the help of quantitative analysis of peripheral immune function, which may inspire performing further investigations on the relevance of immune function with different diseases.

XPG rs873601 G>A contributes to uterine leiomyoma susceptibility in a Southern Chinese population.

XPG gene contributes to DNA repair defects and genomic instability, which may lead to the initiation of uterine leiomyoma. We hypothesized that genetic variants of XPG gene may alter the carriers' susceptibility to leiomyoma. The association between five potential functional single nucleotide polymorphisms (SNPs), i.e. rs2094258 C>T, rs751402 C>T, rs2296147 T>C, rs1047768 T>C, rs873601 G>A, and uterine leiomyoma risk in Chinese, was investigated in this case-control study, which included 398 incident leiomyoma cases and 733 controls. We found that rs873601 was significantly associated with tumor risk in a recessive genetic model after being adjusting for age and menopause. When compared with rs873601 GG/GA genotypes, the AA genotype had an increased leiomyoma risk (adjusted OR = 1.59, 95% CI = 1.16-2.18, P=0.004; Bonferroni adjusted P=0.040). Furthermore, stratified analysis revealed that the association between the rs873601 AA genotype and leiomyoma risk was more evident among subjects younger than 40 years old (adjusted OR = 1.58, 95% CI = 1.06-2.35, P=0.023) and patients who had more than three myomas (adjusted OR = 2.05, 95% CI = 1.24-3.41, P=0.006). Yet, no significant association between the other four polymorphisms and leiomyoma risk was observed. To sum up, the present study reported on the association between XPG gene polymorphisms and myoma risk. The observed data indicated that SNP rs873601 G>A contributes to uterine leiomyoma susceptibility in a Southern Chinese population.