长链非编码RNA XIST、微小RNA-212-3p对人卵巢颗粒细胞增殖和凋亡的影响  

Effects of long non-coding RNA XIST and microRNA-212-3p on proliferation and apoptosis of human ovarian granulosa cells

作　　者：徐潇萌 尹程 董冰 李跃文 刘适 陈隽 XU Xiaomeng;YIN Cheng;DONG Bing;LI Yuewen;LIU Shi;CHEN Juan(Department of Gynecology,the Third Affiliated Hospital of Qiqihar Medical University,Heilongjiang Province,Qiqihar 161000,China)

机构地区：[1]齐齐哈尔医学院附属第三医院妇科,黑龙江齐齐哈尔161000

出　　处：《妇儿健康导刊》2024年第20期182-187,193,共7页JOURNAL OF WOMEN AND CHILDREN'S HEALTH GUIDE

基　　金：黑龙江省齐齐哈尔市科技计划联合引导项目(LSFGG-2023021)。

摘　　要：目的探讨长链非编码RNA(lncRNA)XIST与微小RNA-212-3p(miR-212-3p)对人卵巢颗粒细胞增殖和凋亡的影响。方法采用实时荧光定量PCR检测LncRNAXIST与miR-212-3p在人正常卵巢上皮细胞IOSE80和KGN细胞株中的表达水平;通过Starbase靶基因数据库预测LncRNAXIST靶向miR-212-3p,并采用双萤光素酶报告实验进行验证;分别将XIST-plasmid和miR-212-3pmimic转染至KGN细胞,通过CCK-8实验、流式细胞术和蛋白质印迹法(Westernblot)实验分析LncRNAXIST与miR-212-3p对KGN细胞增殖和凋亡的影响。结果与IOSE80细胞系比较,在KGN细胞中LncRNAXIST呈低表达,miR-212-3p呈高表达(P<0.001);StarBase靶基因数据库预测显示,XIST与miR-212-3p存在结合位点;miR-212-3p mimic可降低XIST-WT的萤光素酶活性(P<0.01);CCK-8实验和流式细胞术实验结果显示,LncRNAXIST能够抑制KGN细胞增殖,促进细胞凋亡,转染miR-212-3pmimic能够部分抑制LncRNAXIST(P<0.001);Western blot实验结果显示,LncRNA XIST能够抑制B淋巴细胞瘤-2蛋白(Bcl-2)表达,促进Bax蛋白的表达,转染miR-212-3p mimic能够部分抑制Bax蛋白表达,促进Bcl-2蛋白的表达(P<0.001)。结论LncRNA XIST过表达能够抑制KGN细胞的增殖并促进其凋亡,其机制作用可能是通过靶向miR-212-3p实现的。Objective To explore the effects of long non-coding RNA(lncRNA)XIST and microRNA-212-3p(miR-212-3p)on the proliferation and apoptosis of human ovarian granulosa cells.Methods The expression levels of LncRNA XIST and miR-212-3p in human normal ovarian epithelial cell lines IOSE80 and KGN were detected by real time fluorogenic quantitative PCR.LncRNA XIST target miR-212-3p was predicted by Starbase target gene database,and verified by luciferase reporter assay.The XIST-plasmid and miR-212-3p mimic were transfected into KGN cells,respectively.The effects of LncRNA XIST and miR-212-3p on the proliferation and apoptosis of KGN cells were analyzed by CCK-8 assay,flow cytometry,and Western blot experiments.Results Compared with IOSE80 cell line,LncRNA XIST expression was low in KGN cells,and miR-212-3p expression was high in KGN cells(P<0.001).The Starbase target gene database predicted that there were binding sites between XIST and miR-212-3p.miR-212-3p mimic reduced the luciferase activity of XIST-WT(P<0.01).The results of CCK-8 assay and flow cytometry experiments showed that LncRNA XIST could inhibit KGN cell proliferation and promote cell apoptosis,and transfection of miR-212-3p mimic could partially inhibit LncRNA XIST(P<0.001).Western blot experiment results showed that LncRNA XIST could inhibit the expression of B-cell lymphoma-2 protein(Bcl-2),and promote the expression of Bax protein.Transfection with miR-212-3p mimic could partially inhibit the expression of Bax protein.It also promoted the expression of Bcl-2 protein(P<0.001).Conclusion LncRNA XIST overexpression can inhibit the proliferation and promote the apoptosis of KGN cells,and its mechanism may be achieved by targeting miR-212-3p.

关 键 词：长链非编码RNA XIST 微小RNA-212-3p 人卵巢上皮细胞 细胞增殖 细胞凋亡 

分 类 号：R711[医药卫生—妇产科学]