miR-375靶向Pax6基因调控视网膜母细胞瘤Y79细胞增殖凋亡的分子机制  

Molecular mechanism of miR-375 targeting Pax6 gene regulating the proliferation and apoptosis of retinoblastoma-Y79 cells

作　　者：桂玉敏 彭建军 郭敬 GUI Yumin;PENG Jianjun;GUO Jing(Department of Ophthalmology,Puren Hospital of Wuhan City,Hubei,Wuhan 430081,China)

机构地区：[1]湖北省武汉市普仁医院眼科,430081 [2]华中科技大学同济医学院附属同济医院眼科

出　　处：《河北医药》2020年第8期1156-1159,共4页Hebei Medical Journal

摘　　要：目的探讨miR-375对人视网膜母细胞瘤细胞增殖和凋亡的影响及作用机制。方法采用脂质体转染法分别将miR-375 mimics或阴性对照NC mimics转染至Y79细胞,采用qRT-PCR检测转染效率,噻唑蓝比色法(MTT)检测miR-375对Y79细胞增殖能力的影响,流式细胞仪检测miR-375对Y79细胞凋亡的影响,生物信息学工具分析miR-375的靶基因,双荧光素酶报告基因实验检测miR-375和Pax6靶向关系,qRT-PCR和Western blot检测miR-375对Pax6的调控作用。结果转染miR-375 mimics后,Y79细胞中miR-375的表达明显升高(P<0.05)。过表达miR-375可抑制Y79细胞的增殖活力(P<0.05),促进Y79细胞凋亡(P<0.05)。生物信息学软件分析结果显示miR-375与Pax63’UTR存在结合位点,双荧光素酶报告基因实验结果显示Pax6是miR-375的靶基因,qRT-PCR和Western blot检测结果显示miR-375可负向调控Pax6 mRNA和蛋白的表达。结论miR-375靶向Pax6基因抑制视网膜母细胞瘤Y79细胞增殖,诱导Y79细胞凋亡。Objective To investigate the effects and action mechanism of miR-375 on proliferation and apoptosis of human retinoblastoma cells.Methods The miR-375 mimics or negative control NC mimics were transfected into Y79 cells by lipofection.The transfection efficiency was detected by qRT-PCR.The efects of miR-375 on the proliferation of Y79 cells were detected by thiazolyl blue colorimetric assay(MTT),and its effects on apoptosis of Y79 cells were detected by flow cytometry.Moreover the bioinformatics tools was used to analyze the target genes of miR-375,dual luciferase reporter gene assay was used to analyze the targeting correlation between miR-375 and Pax6,and qRT-PCR and Western blot were used to detect the regulatory effects of mix-375 on Pax6.Results After transfection of miR-375 mimics,the expression levels of miR-375 in Y79 cells were significantly increased(P<0.05).The overexpression of miR-375 could inhibit the proliferation of Y79 cells and promote the apoptosis of Y79 cells(P<0.05).Bioinformatics software analysis showed that miR-375 and Pax63’UTR had binding sites.The results of dual luciferase reporter gene showed that Pax6 was the target gene of miR-375,and qRT-PCR and Western blot results showed that miR-375 could negatively regulate the expressions of Pax6 mRNA and protein.Conclusion The miR-375 targeting Pax6 gene can inhibit the proliferation of retinoblastoma Y79 cells and induce apoptosis of Y79 cells.

关 键 词：miR-375 PAX6基因 视网膜母细胞瘤Y79细胞 增殖 凋亡 

分 类 号：R774.1[医药卫生—眼科]