miR-143对宫颈癌HeLa细胞增殖和凋亡的影响以及对Bcl-2表达的调节  

Role of miR-143 in proliferation and apoptosis of HeLa cell proliferation and the regulation of Bcl-2

作　　者：刘立鹏[1] 于潇华[1] 王晓春[2] 郭小芳[1] 田智[1] 龙昱[1] 粟敏[1] 罗玥佶[1] 黄春霞[1] 胡桔红[1] 吴新华[3] 宋元达[4] 

机构地区：[1]长沙医学院生化教研室,湖南长沙410219 [2]中南大学湘雅医学院医学检验系 [3]中南大学湘雅医学院妇产科,湖南长沙410013 [4]中南大学生命科学与技术学院生化系,湖南长沙410013

出　　处：《中国药理学通报》2011年第2期265-269,共5页Chinese Pharmacological Bulletin

基　　金：湖南省自然科学基金资助项目(No7JJ3058)

摘　　要：目的研究miR-143对宫颈癌HeLa细胞增殖和凋亡的影响,探讨miR-143对宫颈癌HeLa细胞系Bcl-2蛋白及mRNA表达的调节作用。方法将miR-143,anti-miR-143以及microRNA空对照高表达质粒用脂质体转染进宫颈癌He-La细胞中,MTT法检测宫颈癌HeLa细胞增殖的变化,His-tone/DNA ELISA检测细胞凋亡;Western blot和RT-PCR检测Bcl-2蛋白和mRNA的表达情况;双荧光素酶报告基因法检测miR-143的靶位点。结果 MiR-143抑制HeLa细胞的增殖,促进细胞凋亡,而anti-miR-143促进HeLa细胞增殖,抑制细胞凋亡;高表达的miR-143抑制Bcl-2蛋白的表达,anti-miR-143增加Bcl-2蛋白的表达量,而Bcl-2 mRNA的变化很小;miR-143抑制Bcl-2 3'端的荧光素酶的活性,突变miR-143与Bcl-2的预测结合位点后,荧光素酶的活性恢复。结论 MiR-143至少部分通过打靶Bcl-2抑制宫颈癌HeLa细胞增殖,并且促进HeLa细胞凋亡。Aim To explore the effect of miR-143 on the poliferation and apoptosis of human cervical carcinoma,and investigate the role of miR-143 in the expression of Bcl-2 in HeLa cells.Methods Over-expression plasmids with miR-143,anti-miR-143 and control miRNA precursor were inserted into the HeLa cells using lipofectamine 2000.Methyl thiazolyl tetrazolium（MTT）was used to analyze the proliferation of the HeLa cells,and cell apoptosis was quantitated by Histone/DNA ELISA.The level of Bcl-2 protein and mRNA was measured by using Western blot and RT-PCR respectively.Dual-luciferase reporter gene assay was used to detect the target sites of miR-143.Results miR-143 inhibited significantly HeLa cell proliferation and promoted apoptosis,while anti-miR-143 promoted the cell proliferation,and suppressed the apoptosis.Sustained over-expression of miR-143 resulted in repression of Bcl-2 protein,and anti-miR-143 increased Bcl-2,whereas the levels of Bcl-2 mRNA were so minor.MiR-143 repressed the activity of a luciferase reporter carrying the 3′-untranslated region of Bcl-2,mutated the predicted binding site between miR-143 and Bcl-2,and the activity of luciferase was reversed.Conclusion miR-143 can inhibit proliferation and promote apoptosis in HeLa cells,at least in part by targeting Bcl-2.

关 键 词：MIR-143 BCL-2 增殖 凋亡 HELA细胞 宫颈癌 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]