microRNA-130a抑制KGN细胞FOXL2基因的表达及对细胞增殖的影响

发布时间：2018-02-05 00:56

本文关键词： miR-a 转染 KGN细胞 FOXL 基因表达调控细胞增殖　出处：《中国美容医学》2016年01期 　论文类型：期刊论文

【摘要】：目的:研究外源性microRNA-130a在卵巢颗粒细胞KGN中过表达对FOXL2基因表达的影响,探索microRNA-130a对KGN细胞增殖的影响。方法:人工设计合成靶向FOXL2基因的microRNA-130a模拟物,脂质体法将microRNA-130a模拟物转染入KGN细胞中,Trizol法和RIPA蛋白裂解法分别获取高纯度的细胞总RNA和总蛋白,实时荧光定量聚合酶联反应(FQ-PCR)检测microRNA模拟物转染后细胞中FOXL2基因mRNA表达水平,Western Blot检测FOXL2蛋白表达水平,四甲基偶氮唑盐法(MTT)检测microRNA-130a转染后KGN细胞的增殖情况。结果:成功转染KGN细胞后,阴性对照组FOXL2基因mRNA和蛋白的表达水平与空白对照组相比无明显的差异(P0.05),mi R-130a转染组能显著抑制FOXL2 mRNA和蛋白的表达,与空白对照组和阴性对照组相比均有统计学差异(P0.05)。MTT法检测各组KGN细胞的OD值及细胞增值率,microRNA-130a转染后能明显促进KGN细胞的增殖,与与空白对照组和阴性对照组相比均有统计学差异(P0.05)。结论:外源性的mi R-130a对FOXL2基因mRNA和蛋白表达有明显的抑制作用,且能明显促进KGN细胞的增殖。
[Abstract]:Objective: to study the effect of exogenous microRNA-130a overexpression on FOXL2 gene expression in ovarian granulosa cell KGN and to explore the effect of microRNA-130a on the proliferation of KGN cells. Methods: microRNA-130a mimics targeting FOXL2 gene were designed and synthesized. MicroRNA-130a mimics were transfected into KGN cells by liposome method. The high purity total RNA and total protein were obtained by trizol and RIPA protein cleavage, respectively. The expression level of FOXL2 gene mRNA was detected by real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) and the expression of FOXL2 protein was detected by Western Blot after transfection of microRNA mimics. The proliferation of KGN cells after transfection of microRNA-130a was detected by MTT. Results: after transfection of KGN cells, KGN cells were transfected successfully. The expression level of FOXL2 gene mRNA and protein in the negative control group was not significantly different from that in the blank control group. The expression of FOXL2 mRNA and protein was significantly inhibited in the transfected group. Compared with the blank control group and the negative control group, there were significant differences between the two groups. The OD value of KGN cells and the cell proliferation rate were detected by P0.05. MTT assay. MicroRNA-130a transfection could significantly promote the proliferation of KGN cells. Conclusion: exogenous mi R-130a can significantly inhibit the expression of FOXL2 gene mRNA and protein, and promote the proliferation of KGN cells.
【作者单位】：潍坊医学院整形外科研究所;
【基金】：国家自然科学基金(81272122 81301647 81471880) 山东省科技攻关计划(JK67) 山东省自然科学基金(ZR2013HQ025)
【分类号】：Q78;R329.2
【正文快照】： FOXL2是叉头框转录因子家族成员之一,主要表达在眼周、卵巢和垂体细胞中,该基因最早由Crisponi作为小睑裂综合征(BPES,Blepharophimosis Syndrome,MIM:110100)及卵巢早衰(premature ovarian failure,POF)的候选基因被克隆定位[1]。小睑裂综合征是一种常染色体显性遗传病[2],临

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