lncRNA H19 acts as a ceRNA to regulate the expression of CTGF by targeting miR-19b in polycystic ovary syndrome

Abstract

The etiology of polycystic ovary syndrome (PCOS) is complex and the pathogenesis is not fully understood. Some studies have shown that dysregulation of ovarian granulosa cells may be related to abnormal follicles and excessive androgen in women with PCOS. Our team has also confirmed the high expression status of H19 in PCOS patients in the early stage. However, the relationship between H19 and miR-19b in the development of PCOS is still unknown. Therefore, we used bioinformatics to predict the binding sites of human H19 and miR-19b, and of miR-19b and CTGF genes. After the silencing and overexpression of H19, real-time polymerase chain reaction (PCR) was used to detect the expressions of H19, miR-19b, and CTGF. Western blotting was used to detect CTGF protein. Proliferation of KGN cells after H19 silencing was detected by CCK8. Flow cytometry was used to detect the apoptosis of KGN cells after H19 silencing. After the overexpression of H19, it was found that the expression of miR-19b gene decreased and the expression of CTGF increased, whereas silencing of H19 did the opposite. In addition, H19 could promote cell proliferation and decrease cell apoptosis. Finally, luciferase reporter assays showed that the 3 0-end sequences of lncRNA H19 and CTGF contained the binding site of miR-19b. In conclusion, our study indicated that lncRNA H19 acted as a ceRNA to bind to miR-19b via a ''sponge'' to regulate the effect of CTGF on KGN cells, which may play a vital role in PCOS.

microRNAs and long non‐coding RNAs as biomarkers for polycystic ovary syndrome

“…Yang et al 95 LncRNA-BANCR -Role in PCOS by promoting apoptosis in granulosa cells -From cells of IVF patients Wang et al 54 LncRNA-GAS5 -LncRNA-GAS5 is upregulated in polycystic ovary syndrome and regulates cell apoptosis and the expression of IL−6 in granulosa cells -From blood plasma Sun et al 53 lncRNA-H19 -lncRNA H19 acts as a ceRNA to regulate the expression of CTGF by targeting miR−19b in PCOS -On KGN cell line -H19 could promote cell proliferation and decrease cell apoptosis Chen et al 96 LncRNA-HCP5 -LncRNA-HCP5 promotes cell proliferation and inhibits apoptosis via miR−27a−3p/ IGF−1 axis -On human granulosa-like tumor cell line KGN Guo et al 97 LncRNA-HOTAIRM1 -LncRNA-HOTAIRM1/miR−433-5p/PIK3CD function as a ceRNA network to encourage the development of PCOS 101 LncRNA-H19 -LncRNA-H19 is associated with PCOS in Chinese women -From peripheral blood leukocytes Huang et al 47 Lnc-CCNL1-3:1 -lnc-CCNL1-3:1 promotes granulosa cell apoptosis and suppresses glucose uptake in PCOS -From human luteinized granulosa cells(hLGCs) derived from women Zhao et al 102 LINC−01572:28 -LINC−01572:28 inhibits granulosa cell growth via a decrease in p27 in PCOS Sang et al 103 LncRNA-NEAT1 -LncRNA-NEAT1 drives the development of PCOS via sponging multiple miRNAs Youssef et al 104 LncRNA steroid receptor activator (SRA)…”

“…Also, in order to trigger the attention on new biomarkers for the disease, studies on cell lines can also be favorable, especially when the line is correlated to a special condition of the disease, like that for KGN as performed by many groups. 17,31,44,47,53,54 Blood serum seems feasible for clinical diagnostics, and statistically significant biomarkers in the blood are not showed that oocyte lncRNAs could have roles in chromatin remodeling, cell pluripotency and also in the early development of the embryo. They did so by analyzing the differential lncRNA expression profiles in human oocytes and cumulus cells.…”

“…Also, in order to trigger the attention on new biomarkers for the disease, studies on cell lines can also be favorable, especially when the line is correlated to a special condition of the disease, like that for KGN as performed by many groups. 17 , 31 , 44 , 47 , 53 , 54 Blood serum seems feasible for clinical diagnostics, and statistically significant biomarkers in the blood are not few. It is important to note that a differentially present biomolecule in the blood components is a very significant biomarker for the syndrome and not necessarily an early stage indicator.…”

Non‐coding RNAs and their cross‐talks impacting reproductive health of women

“…Lately, a study reported a well‐characterized lncRNA H19 acting as a ceRNA in human ovarian granulosa KGN cells by competitively binding to miR‐19b and regulating the expression of CTGF. The study revealed that elevated expression of CTGF promotes GC proliferation while inhibiting apoptosis, thereby promoting KGN cell proliferation (Sun et al, 2020). Taken together, this study provided evidence that aberrantly expressed lncRNA H19 in GC plays a pivotal role as a ceRNA by sponging miRNAs, which might underlie the pathophysiology of PCOS.…”

(In)Distinctive Role of Long Non-Coding RNAs in Common and Rare Ovarian Cancers

“…Benign ovarian teratomas show a varying degree of H19 hypomethylation, and DNA prepared from cultured teratoma cells shows extreme hypomethylation of the H19 locus [154]. Ovarian granulosa cell tumor cell line KGN has been used to demonstrate that lncRNA H19 binds to miR-19b, resulting in upregulation of CTFG, increase in cell proliferation, d reduction in the rate of apoptosis in these cells [155].…”