乳腺癌扩增序列2基因RNA干扰慢病毒载体的构建及对乳腺癌MDA-MB-231细胞增殖的影响  

Construction of breast cancer amplified sequence 2 gene lentiviral RNA interference vector and effect on proliferation of human breast cancer cells MDA-MB-231

作　　者：石杰 白腾飞 陈悦 麦若玫 韩翠翠[1] 林宇[1] SHI Jie;BAI Teng-fei;CHEN Yue;MAI Ruo-mei;HAN Cui-cui;LIN Yu(School of Pharmacy,Qiqihar Medical University,Qiqihar 161006,Heilongjiang Province,China)

机构地区：[1]齐齐哈尔医学院药学院,黑龙江齐齐哈尔161006

出　　处：《中国临床药理学杂志》2020年第7期789-791,795,共4页The Chinese Journal of Clinical Pharmacology

基　　金：黑龙江省自然科学基金面上基金资助项目(C2016060);黑龙江省大学生创新创业训练计划基金资助项目(201711230016);齐齐哈尔医学院博士基金资助项目(QY2016B-24)。

摘　　要：目的构建乳腺癌扩增序列2(BCAS2)基因RNA干扰(RNAi)的细胞模型,研究BCAS2对乳腺癌MDA-MB-231细胞增殖的影响。方法将人乳腺癌MDA-MB-231细胞分为5组:空白组、对照组、3个干扰组。空白组细胞不做任何处理,对照组细胞转染对照慢病毒载体,干扰组细胞转染慢病毒载体pGLV3-GFP-shBCAS2(shBCAS2-1,2,3)。用实时荧光定量-PCR法和蛋白质印迹法分别检测转染后MDA-MB-231细胞中BCAS2基因及其蛋白表达水平,MTT法检测shBCAS2对MDA-MB-231细胞增殖能力(光密度值)的影响。结果空白组、对照组和3个干扰组的BCAS2基因表达量分别为1.00±0.07,0.99±0.08,0.21±0.06,0.48±0.13和0.37±0.12;这5组的蛋白相对表达量分别为1.32±0.04,1.33±0.04,0.23±0.06,0.54±0.09和0.77±0.07;干扰组与对照组相比,BCAS2基因与蛋白表达水平均明显下降,差异均有统计学意义(均P<0.05)。第3天,干扰shBCAS2-1组和对照组的细胞增殖能力分别为0.70±0.04和0.84±0.02,组间比较差异有统计学意义(P<0.01)。结论 shRNA干扰BCAS2可抑制乳腺癌MDA-MB-231细胞的增殖。Objective To establish a cell model of RNA interference(RNAi) of the breast cancer amplification sequence 2(BCAS2) gene, and study the effect of BCAS2 on the proliferation of breast cancer MDA-MB-231 cells.Methods Human breast cancer MDA-MB-231 cells were divided into five groups: blank group, control group, interference group 1, interference group 2 and interference group 3.No treatment was taken in blank group, and the control lentivirus vector was transfected in control group, while the different lentivirus vectors pGLV3-GFP-shBCAS2(shBCAS2-1, 2, 3) were transfected in different interference groups,respectively.After transfected, mRNA and protein expression of BCAS2 in MDA-MB-231 cells were detected by real-time quantitative-PCR and Western blot,respectively.The effect of shBCAS2 on the proliferation(OD value) of MDA-MB-231 cells was determined by MTT assay.Results The mRNA expressions of BCAS2 in blank group, control group and three interference groups were 1.00±0.07,0.99±0.08,0.21±0.06,0.48±0.13 and 0.37±0.12,respectively;the relative protein expression of BCAS2 in the five groups were 1.32±0.04,1.33±0.04,0.23±0.06,0.54±0.09 and 0.77±0.07,respectively.Compared with the control group,the mRNA and protein expression levels of BCAS2 in the interference groups all decreased significantly(all P < 0.05).The cell proliferation rate in shBCAS2-1 group and control group were0.70±0.04,0.84±0.02 at third day,and comparing between the two groups,the difference was significant(P < 0.01).Conclusion The proliferation of breast cancer MDA-MB-231 cells can be inhibited by shRNA interference with BCAS2.

关 键 词：RNA干扰 乳腺癌扩增序列2 慢病毒表达载体 乳腺癌 细胞增殖 

分 类 号：R97[医药卫生—药品]