微小RNA 761对人卵巢颗粒细胞增殖和凋亡的影响

Effects of microRNA 761 on proliferation and apoptosis of human ovarian granulocyte cells

目的研究微小RNA 761(miR-761)对人卵巢颗粒细胞增殖和凋亡的影响及其机制。方法取多囊卵巢综合征(PCOS)患者卵巢颗粒细胞培养,随机分为3组,分别为miR-761 inhibitor组(转染miR-761抑制剂),miR-761 mimic组(转染miR-761模拟物),miR-NC组(转染阴性对照寡核苷酸)。用细胞计数法-8(CCK-8)实验检测转染细胞的增殖能力;用TUNEL实验检测转染细胞的凋亡能力;用蛋白质印迹(Western blot)法检测雌激素受体2(estrogen receptor 2,ESR2)的表达。采用双荧光素酶报告基因实验验证miR-761与ESR2之间的结合关系。结果miR-NC组、miR-761 mimic组和miR-761 inhibitor组的细胞存活率分别为(98.33±1.53)%,(121.23±3.74)%,(79.86±3.66)%;细胞凋亡率分别为(6.51±0.26)%,(4.41±0.24)%和(10.48±0.33)%;ESR2蛋白表达水平分别为1.05±0.03,0.74±0.08和3.90±0.28。以上指标,miR-NC组和miR-761 mimic组相比,miR-NC组和miR-761 inhibitor组相比,差异均有统计学意义(均P<0.05)。双荧光素酶报告基因实验证实miR-761可与ESR2靶向结合。结论miR-761通过下调ESR2促进PCOS患者卵巢颗粒细胞增殖并抑制细胞凋亡。

Objective To investigate the effects of microRNA-761(miR-761)on the proliferation and apoptosis of human ovarian granular cells and its mechanism.Methods The ovarian granular cells of polycystic ovary syndrome(PCOS)patients were cultured and randomly divided into three groups:miR-761 inhibitor group(transfected with miR-761 inhibitors),miR-761 mimic group(transfected with miR-761 mimics),and miR-NC group(transfected with negative control oligonucleotides).Cell counting kit-8(CCK-8)assay was used to detect the proliferation ability of the transfected cells.TUNEL assay was used to detect the apoptosis of the transfected cells.The expression of estrogen receptor 2(ESR2)was examined by Western blot assay.The binding relationship between miR-761 and ESR2 was verified by dual-luciferase reporter gene assay.Results The cell survival rates of the miR-NC group,miR-761 mimic group and miR-761-inhibitor group were(98.33±1.53)%,(121.23±3.74)%and(79.86±3.66)%,respectively;the apoptosis rates were(6.51±0.26)%,(4.41±0.24)%and(10.48±0.33)%,respectively;the ESR2 protein expression levels were 1.05±0.03,0.74±0.08 and 3.90±0.28,respectively.In terms of the above indicators,the differences were statistically significant between miR-NC group and miR-761 mimic group,as well as between miR-NC group and miR-761 inhibitor group(all P<0.05).Dual-luciferase reporter gene assay confirmed that miR-761 could target and bind to ESR2.Conclusion MiR-761 promotes the proliferation and inhibits the apoptosis of ovarian granular cells of PCOS patients by down-regulating the expression of ESR2.