Mechanism of LncRNA SRA Regulating Chronic Inflammation in KGN through MEK/ERK/GATA4 Pathway
XU Fang1, LIANG Jinling2, PAN Zimeng2, MA Ming3, SUN Miao1*, KUANG Hongying1*
The research purpose of this article was to explore the regulatory mechanism of LncRNA SRA in PCOS ovarian local inflammation and the possible role of MEK/ERK/GATA4 pathway in this regulatory mechanism. Human ovarian granulosa cell tumor cell line (KGN) was selected in this study. KGN was divided into nontransfected group (Control), LncRNA SRA overexpression control group (Vector), LncRNA SRA overexpression vector transfection group (LncRNA SRA), LncRNA SRA overexpression Vector+PD98059 treatment group (inhibitor of MEK) (Vector+PD98059), and LncRNA SRA overexpression vector+PD98059 treatment group (LncRNA SRA+PD98059) by cell transfection technology. RT-qPCR was used to detect the expression level of LncRNA SRA in each group. MTT assay was used to detect cell proliferation. Cell apoptosis was detected by flow cytometry. The protein expression levels of cleaved-Caspase-3, Caspase-3, p-MEK1/2, MEK1/2, p-ERK1/2, ERK1/2, p-GATA4, GATA4, p-p65 and SGMS2 were detected by Western blot. The expression levels of TNF-α, IL-6 and IL-1β were detected by ELISA. The results showed that inhibition of MER pathway could not only reduce the increased proliferation and inflammation of KGN cells caused by LncRNA SRA and the high expression level of MEK/ERK/GATA4 pathway related proteins, but also increase the decreased apoptosis of KGN cells caused by LncRNA SRA. These results suggest that LncRNA SRA/MEK/ERK/GATA4 pathway may play an important role in regulating the local inflammatory microenvironment of PCOS ovarian granular cells.
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