MicroRNA-93 Promotes Ovarian Granulosa Cells Proliferation Through Targeting CDKN1A in Polycystic Ovarian Syndrome
MicroRNA-93 Promotes Ovarian Granulosa Cells Proliferation Through Targeting CDKN1A in Polycystic Ovarian Syndrome
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MicroRNA-93 通过靶向多囊卵巢综合征中的 CDKN1A 促进卵巢颗粒细胞增殖
DOI：
10.1210/jc.2014-3827
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发表时间：
2015-05-01
影响因子：
5.8
通讯作者：
Yang, Dongzi
中科院分区：
文献类型：
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作者：
Jiang, Linlin;Huang, Jia;Yang, Dongzi
背景：MicroRNAs （miRNAs）是一种小的非编码rna，在转录后负调控基因表达。不同表达的miRNAs是否有助于促进多囊卵巢综合征（PCOS）患者颗粒细胞增殖尚不清楚。目的：探讨某些mirna是否参与PCOS卵巢功能障碍及颗粒细胞增殖增加的机制。患者和细胞：分析了16例PCOS患者和8例非PCOS患者切除卵巢皮质中miRNA的表达。用永生化人颗粒细胞（KGN）进行了机制研究。主要观察指标：使用qRT-PCR检测卵巢皮质mirna的表达，并在miR-93过表达或抑制或胰岛素治疗后培养KGN颗粒细胞进行增殖试验。利用生物信息学技术鉴定潜在的miRNA靶点。采用蛋白表达分析、荧光素酶测定和挽救测定来确认miR-93的底物。结果：MiR-93在PCOS卵巢皮质中表达升高，其鉴定的靶点CDKN1A下调。MiR-93过表达促进细胞增殖和G1向S过渡。敲低CDKN1A促进颗粒细胞的细胞生长和细胞周期进程，CDKN1A的重新引入逆转了miR-93的促进作用。高浓度胰岛素诱导miR-93上调，刺激KGN细胞增殖，降低CDKN1A表达。结论：miR-93在PCOS颗粒细胞中升高，靶向CDKN1A促进增殖和细胞周期进展。胰岛素可上调miR-93的表达，促进细胞增殖。这可能为多囊卵巢综合征颗粒细胞功能障碍提供新的认识。
Context: MicroRNAs (miRNAs) are small, noncoding RNAs that negatively regulate gene expression post-transcriptionally. Whether differently expressed miRNAs contribute to promoting granulosa cell proliferation in polycystic ovarian syndrome disease (PCOS) remains unknown.Objective: We explored whether certain miRNAs are involved in the ovarian dysfunction of PCOS and the mechanism of increased granulosa cells proliferation.Patients and Cells: miRNA expression was analyzed in excised ovarian cortexes from 16 women with PCOS and 8 non-PCOS. An immortalized human granulosa (KGN) cell was used for the mechanism study.Main Outcome Measures: Expressions of miRNAs in ovarian cortexes were measured using qRT-PCR and KGN granulosa cells were cultured for proliferation assays after overexpression or inhibition of miR-93 or after insulin treatment. Bioinformatics were used to identify the potential miRNA targets. Protein expression analysis, luciferase assays, and rescue assays were used to confirm the substrate of miR-93.Results: MiR-93 expression was higher in PCOS ovarian cortex and its identified target, CDKN1A, was downregulated. MiR-93 overexpression promoted cell proliferation and G1 to S transition. Knocking down CDKN1A promoted cell growth and cell cycle progression in granulosa cells, and CDKN1A re-introduction reversed the promotional role of miR-93. High concentrations of insulin induced upregulation of miR-93, stimulated KGN cells proliferation and reduced CDKN1A expression.Conclusions: miR-93 was increased in PCOS granulosa cells and targeted CDKN1A to promote proliferation and cell cycle progression. Insulin could upregulate the expression of miR-93 and stimulate cell proliferation. This might provide a new insight into the dysfunction of granulosa cells in PCOS.