LncRNA LNC-GULP1-2:1 is involved in human granulosa cell proliferation by regulating COL3A1 gene

Abstract

Long non-coding RNAs (lncRNAs) are defined as transcripts longer than 200 nucleotides that are not translated into protein. Many studies have identified the association between abnormal expression of lncRNAs and specific disease states, especially in tumor-related studies. However, the precise functions of these lncRNAs are not clear. In our previous study, both lnc-GULP1-2:1 and its potential target COL3A1 were significantly downregulated in ovarian cortical tissues of POI patients compared with normal control patients. The results of GO and KEGG analysis also showed that changes in extracellular matrix-related genes play an important role in the development of POI. Therefore, we speculate that lnc-GULP1-2:1 may participate in the development of ovarian follicles by regulating the COL3A1 gene. Basic research study. The lentiviral vectors were used to construct a KGN cell line with stable high expression of lnc-GULP1-2:1. Real-time PCR and western blot were used to analyze the expression of COL3A1 and lnc-GULP1-2:1. Cell proliferation was analyzed by using CCK-8 kits. The stable over-expression of lnc-GULP1-2:1 in KGN cells could significantly inhibit granulosa cell proliferation. Treatment of KGN cells with chemotherapeutic agents (cisplatin and paclitaxel) dose-dependently inhibited granulosa cell proliferation and promoted the expression of lnc-GULP1-2:1, whereas the cytokine TGFα, which promoted granulosa cell proliferation, inhibited the expression of lnc-GULP1- 2:1. These results suggest that lnc-GULP1-2:1 may be involved in proliferation regulation of granulosa cells. Further studies found that the protein levels of COL3A1 were significantly increased in KGN cells with stable over-expression of lnc-GULP1-2:1, indicating that lnc-GULP1-2:1 regulates the expression of COL3A1. Levels of lnc-GULP1-2:1 can be down-regulated by siRNAs (5 nM, 10 nM , and 20 nM) in different concentrations of COL3A1 in KGN cells, and silencing COL3A1 could also significantly inhibit KGN cell proliferation. The results of this study suggest that the involvement of lnc-GULP1-2:1 in the regulation of KGN granulosa cells may partly be mediated through the regulation of its target gene COL3A1. The regulation effect of lnc-GULP1-2:1 on granulosa cell proliferation was consistent with the occurrence of POI in clinical chemotherapeutic agents, suggesting that lnc-GULP1-2:1 may be involved in the regulation of granulosa cell function in POI patients. This is the first study to discover the regulation of lnc-GULP1-2:1 in ovarian granulosa cells, which is of great significance for the further investigation of lncRNA regulation in ovarian follicle development.