靶向作用TIMP-2调控人卵巢颗粒细胞的增殖能力

doi:10.12092/j.issn.1009-2501.2020.05.007

摘要/Abstract

摘要： 目的:探讨miR-106a对人卵巢颗粒细胞(KGN)增殖能力的调节作用,并初步探讨其可能的作用靶点。方法:采用细胞转染法调控miR-106a在颗粒细胞中的表达,RT-PCR法检测其表达水平。采用MTT法检测细胞的增殖活性,生物信息学预测miR-106a可能的靶基因并采用双荧光素酶实验验证,Western blot法检测靶蛋白的表达。结果:miR-106a在KGN细胞中表达显著高于正常卵巢上皮细胞(IOSE80),差异有统计学意义(P<0.05),抑制miR-106a表达后KGN细胞的增殖活性显著降低(P<0.05),双荧光素酶实验结果证实miR-106a可直接靶向作用于TIMP-2基因,Western blot结果显示过表达miR-106a后TIMP-2蛋白的表达水平显著降低(P<0.05)。结论:miR-106a可促进KGN细胞的增殖,与靶向降低TIMP-2表达水平有关。

关键词: miR-106a, TIMP-2, 增殖, 卵巢颗粒细胞, 多囊卵巢综合征

Abstract: AIM: To investigate the regulation function of miR-106a on the proliferation of human ovarian granulosa cells, and to explore its possible target. METHODS: The expression of miR-106a in granulosa cells was regulated by cell transfection, and its expression level was detected by RT-PCR. The MTT assay was used to detect the cell proliferation activities of cells. Bioinformatics methods were used to predicted the possible target genes of miR-106a, which were verified them by double-luciferase assay. The expression of target protein was detected by Western blot. RESULTS: The expression of miR-106a in KGN cells was significantly higher than that of normal ovarian epithelial cell (IOSE80), the difference was statistically significant (P<0.05). The proliferation activity of KGN cells was significantly decreased after inhibiting the expression of miR-106a (P<0.05). The results of dual luciferase assay showed that miR-106a could directly target TIMP-2 gene. Western blot results showed that the expression level of TIMP-2 protein was significantly decreased after overexpression of miR-106a (P<0.05). CONCLUSION: miR-106a can promote the proliferation of KGN cell; The mechanism is related to the targeted reduction of TIMP-2 expression level.

Key words: miR-106a, TIMP-2, proliferation, ovarian granulosa cells, polycystic ovary syndrome

中图分类号:

R965.2

欧珺, 侯文文, 唐静文, 栗家平, 徐庆阳. 靶向作用TIMP-2调控人卵巢颗粒细胞的增殖能力[J]. 中国临床药理学与治疗学, 2020, 25(5): 527-532.

OU Jun, HOU Wenwen, TANG Jingwen, LI Jiaping, XU Qingyang. miR-106a regulates the proliferation of ovarian granulosa cells by targeting TIMP-2[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2020, 25(5): 527-532.

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