miR-16对人乳腺癌细胞MDA-MB-231增殖和凋亡的作用及机制  

Effect and mechanism of miR-16 on proliferation and apoptosis of human breast cancer cells MDA-MB-231

作　　者：刘嘉祺[1] 周福波[1] 林峰[1] 付惠[1] LIU Jiaqi;ZHOU Fubo;LIN Feng;FU Hui(Department of Pharmacology,Mudanjiang Medical University,Heilongjiang Province,Mudanjiang 157011,China)

机构地区：[1]牡丹江医学院药理教研室,黑龙江牡丹江157011

出　　处：《中国医药导报》2023年第25期38-42,共5页China Medical Herald

基　　金：黑龙江省省属高校基本科研业务费项目(2018-KYYWF-0144)。

摘　　要：目的探究miR-16对人乳腺癌细胞MDA-MB-231增殖和凋亡的作用及机制。方法体外培养细胞,分组如下:人乳腺上皮细胞HBL-100(HBL-100组)和人乳腺癌细胞MDA-MB-231(MDA-MB-231组)。采用荧光实时定量PCR检测miR-16表达,蛋白质印迹法检测β2肾上腺素受体(β2-AR)蛋白表达。细胞瞬时转染,分组如下:MDA-MB-231组,MDA-MB-231+miR-16拟似物(miR-16 mimics组),MDA-MB-231+miR-16抑制物(miR-16 inhibitor组),MDA-MB-231+阴性对照(阴性对照组)。MTT比色法和Annexin V-FITC双染流式细胞仪检测观察miR-16对MDA-MB-231细胞增殖和凋亡影响。双萤光素酶报告基因验证miR-16与β2-AR靶向关系。结果MDA-MB-231组miR-16表达低于HBL-100组,β2-AR蛋白表达高于HBL-100组,差异有统计学意义(P<0.05)。MDA-MB-231组与阴性对照组细胞增殖比较,差异无统计学意义(P>0.05)。miR-16 mimics组细胞增殖低于MDA-MB-231组,miR-16 inhibitor组细胞增殖高于MDA-MB-231组,差异有统计学意义(P<0.05)。MDA-MB-231组与阴性对照组细胞凋亡率比较,差异无统计学意义(P>0.05)。miR-16 mimics组细胞凋亡率高于MDA-MB-231组,miR-16 inhibitor组细胞凋亡率低于MDA-MB-231组,差异有高度统计学意义(P<0.01)。双萤光素酶报告基因结果显示,与MDA-MB-231组比较,miR-16 inhibitor组和阴性对照组萤光素酶活性都没有明显变化,差异无统计学意义(P>0.05)。与MDA-MB-231组比较,具有β2-AR的3’UTR序列的miR-16 mimics组细胞的萤光素酶活性降低,差异有高度统计学意义(P<0.01);与MDA-MB-231组比较,具有突变的β2-AR的3’UTR序列的miR-16 mimics组细胞萤光素酶活性无明显改变,差异无统计学意义(P>0.05)。结论miR-16可能通过靶向β2-AR调控乳腺癌细胞增殖与凋亡。Objective To investigate the effect and mechanism of miR-16 on proliferation and apoptosis of human breast cancer cells MDA-MB-231.Methods The cells cultured in vitro were divided into the following groups:human mammary epithelial cells HBL-100(HBL-100 group)and human breast cancer cells MDA-MB-231(MDA-MB-231 group).Real-time fluorescent quantitative PCR was used to detect miR-16 expression,and Western blot was used to detectβ2 adrenergic receptor(β2-AR)protein expression.Transient transfection of cells was grouped as follows:MDA-MB-231 group,MDA-MB-231+miR-16 mimics(miR-16 mimics group),MDA-MB-231+miR-16 inhibitor(miR-16 inhibitor group),MDA-MB-231+negative control(negative control group).The effects of miR-16 on proliferation and apoptosis of MDA-MB-231 cells were observed by MTT colorimetry and Annexin V-FITC double dye flow cytometry.The targeting relationship between miR-16 andβ2-AR was verified by dual luciferase reporter genes.Results The expression of miR-16 in MDA-MB-231 group was lower than that in HBL-100 group,and the expression ofβ2-AR protein was higher than that in HBL-100 group,and the differences were statistically significant(P<0.05).There was no significant difference in cell proliferation between MDA-MB-231 group and negative control group(P>0.05).Cell proliferation in miR-16 mimics group was lower than that in MDA-MB-231 group,and cell proliferation in miR-16 inhibitor group was higher than that in MDA-MB-231 group,and the differences were statistically significant(P<0.05).There was no significant difference in apoptosis rate between MDA-MB-231 group and negative control group(P>0.05).The apoptosis rate in miR-16 mimics group was higher than that in MDA-MB-231 group,and the apoptosis rate in miR-16 inhibitor group was lower than that in MDA-MB-231 group,and the differences were highly statistically significant(P<0.01).The results of dual luciferase reporter gene showed that,compared with MDA-MB-231 group,luciferase activity in miR-16 inhibitor group and negative control group had no signi

关 键 词：miR-16 Β2肾上腺素受体 MDA-MB-231 增殖 凋亡 

分 类 号：R730.2[医药卫生—肿瘤]