miR-195调控人乳腺癌细胞MCF-7增殖与凋亡的作用及机制研究  

Effect and mechanism of miR-195 regulates on proliferation and apoptosis of human breast cancer MCF-7 cells

作　　者：刘嘉祺[1] 翟凤国[1] 高金霞[2] 刘佳维[3] 杨旭东[1] 周福波[1] 李孟全 LIU Jiaqi;ZHAI Fengguo;GAO Jinxia;LIU Jiawei;YANG Xudong;ZHOU Fubo;LI Mengquan(Department of Pharmacology,Mudanjiang Medical University,Heilongjiang Province,Mudanjiang157011,China;Department of Preventive Medicine,Mudanjiang Medical University,Heilongjiang Province,Mudanjiang157011,China;Department of Organic Chemistry,Mudanjiang Medical University,Heilongjiang Province,Mudanjiang157011,China;Medical Research Center,Mudanjiang Medical University,Heilongjiang Province,Mudanjiang157011,China)

机构地区：[1]牡丹江医学院药理教研室,黑龙江牡丹江157011 [2]牡丹江医学院预防医学教研室,黑龙江牡丹江157011 [3]牡丹江医学院有机化学教研室,黑龙江牡丹江157011 [4]牡丹江医学院医药研究中心,黑龙江牡丹江157011

出　　处：《中国医药导报》2022年第1期18-22,共5页China Medical Herald

基　　金：黑龙江省卫生计生委科研课题(2018487)。

摘　　要：目的研究miR-195调控人乳腺癌细胞MCF-7(MCF-7)增殖与凋亡作用及机制。方法体外培养正常乳腺上皮细胞MCF-10A(MCF-10A)和MCF-7,qRT-PCR检测miR-195表达,蛋白质免疫印迹(WB)检测蛋白磷酸酶1D(PPM1D)蛋白表达。MCF-7瞬时转染,3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法、DNA断裂原位末端标记法观察miR-195对MCF-7增殖和凋亡的影响。双荧光素酶报告基因验证miR-195与PPM1D靶向的关系。结果MCF-7组miR-195表达低于MCF-10A组,PPM1D蛋白表达高于MCF-10A组,差异有统计学意义(P<0.05)。miR-195 mimics组细胞活力低于MCF-7组,miR-195 inhabitor组细胞活力高于MCF-7组,差异有高度统计学意义(P<0.01)。miR-195 mimics组细胞凋亡率高于MCF-7组,差异有高度统计学意义(P<0.01);miR-195 inhabitor组细胞凋亡率低于MCF-7组,差异有高度统计学意义(P<0.05)。双荧光素酶报告基因结果显示,含有PPM1D基因片段miR-195 mimics组荧光素酶活性低于MCF-7组,差异有高度统计学意义(P<0.01);含有突变PPM1D基因片段miR-195 mimics组与MCF-7两组荧光素酶活性比较,差异无统计学意义(P>0.05)。结论miR-195可能通过靶向PPM1D调控乳腺癌细胞增殖与凋亡。Objective To explore the role and mechanism of miR-195 in regulating proliferation and apoptosis of human breast cancer MCF-7 cells(MCF-7).Methods Breast epithelial cells MCF-10A(MCF-10A)and MCF-7 were cultured in vitro.The expression of miR-195 was detected by qRT-PCR.Protein expression of phosphatase magnesium-dependent 1 delta(PPM1D)was detected by Western blot(WB).Transient transfection of MCF-7 was carried out.3-(4,5-two methyl thiazole-2)-2,5-two phenyl tetrazolium bromide(MTT)method and DNA breakage TdT-mediated-duTP nick end labeling method were used to observe the effect of miR-195 on proliferation and apoptosis of MCF-7.Double luciferase reporter genes were used to verify the relationship between miR-195 and PPM1D.Results The expression of miR-195 in MCF-7 group was lower than that in MCF-10A group,the expression of PPM1D protein in MCF-7 group was higher than that in MCF-10A group,the differences were statistically significant(P<0.05).The cell viability of miR-195 mimics group was lower than MCF-7 group;miR-195 inhabitor group was higher than MCF-7 group,the differences were highly statistically significant(P<0.01).The apoptosis rate of miR-195 mimics group was higher than MCF-7 group,the difference was highly statistically significant(P<0.01);and miR-195 inhabitor group was lower than MCF-7 group,the difference was statistically significant(P<0.05).Double luciferase reporter gene showed that miR-195 mimics group,which carried the PPM1D gene fragment,the activity of luciferase was lower than MCF-7 group , the difference was statistically significant (P < 0.01). There was no significant difference in luciferase activity between the mir-195 MIMics group containing the mutated PPM1D gene fragment and McF-7 group (P > 0.05). Conclusion miR-195 may regulate the proliferation and apoptosis of breast cancer cells through targeting PPM1D.

关 键 词：miR-195 蛋白磷酸酶1D 人乳腺癌细胞MCF-7 增殖 凋亡 

分 类 号：R730.2[医药卫生—肿瘤]