AIB1基因沉默对三阴性乳腺癌细胞生长抑制和凋亡的影响  

Growth inhibition and apoptosis induction of AIB1 gene in triple-negative breast cancer cells in vitro

作　　者：王波 董小林 WANG Bo;DONG Xiaolin(Department of Organ Transplantation and Breast Surgery,Affiliated Dongfeng Hospital,Hubei University of Medicine,Shiyan Hubei 442000,China;Department of Physical Examination Center,Affiliated People’s Hospital,Hubei University of Medicine,Shiyan Hubei 442000,China)

机构地区：[1]湖北医药学院附属东风医院器官移植乳腺外科,湖北十堰442000 [2]湖北医药学院附属人民医院健康管理中心,湖北十堰442000

出　　处：《临床与病理杂志》2020年第5期1093-1099,共7页Journal of Clinical and Pathological Research

摘　　要：目的:分析RNAi介导的乳腺癌扩增基因1(amplified in breast cancer 1,AIB1)基因沉默对三阴性乳腺癌细胞生长抑制和凋亡的影响。方法:设计、合成沉默AIB1基因的特异shRNA(短发卡RNA),构建重组表达质粒,转染三阴性乳腺癌细胞MDA-MB-231,蛋白质印迹法检测转染后AIB1蛋白质表达的变化;MTT法检测细胞的生长抑制情况,流式细胞仪检测细胞的凋亡率。结果:本实验成功构建了靶向AIB1基因的RNAi重组表达质粒,并稳定转染三阴性乳腺癌细胞MDA-MB-231。蛋白质印迹法结果表明:稳定转染组AIB1基因的蛋白质表达抑制率为70%,与未转染组相比差异有统计学意义(P<0.05);AIB1基因沉默三阴性乳腺癌细胞24,48,72 h的OD值分别为24.363±0.501,32.596±0.348及43.408±0.257,凋亡率分别为(8.713±0.106)%,(11.396±0.147)%及(14.110±0.117)%,OD值各组间差异有统计学意义(F=43.436,P<0.001);凋亡率各组间差异有统计学意义(F=79.751,P<0.001)。结论:AIB1基因沉默可明显抑制三阴性乳腺癌MDA-MB-231细胞的生长和促进凋亡。Objective:To investigate the silencing effects of amplified in breast cancer 1(AIB1)gene via RNAi on the proliferation and apoptosis of triple-negative breast cancer cell line MDA-MB-231.Methods:According to cDNA sequence of the AIB1 gene,the shRNA which specified silencing AIB1 gene was designed and synthesized,then transfected into MDA-MB-231 cells.The expression levels of AIB1 gene was detected by Western blotting analysis after transfection.MTT assay was applied to evaluate the cell proliferation,flow cytometry assay was conducted to estimate cell apoptosis after being transfected.Results:Our experiments successfully built a recombinant expression plasmid which specified targeting AIB1 gene via RNAi in MDA-MB-231 cells.After transfecting,the protein relative levels of AIB1 in AIB1 gene silencing group were significantly lower as compared with the blank group(P<0.05).The OD value and apoptosis rate in AIB1 gene silencing group after 24,48 and 72 h were 24.363±0.501,32.596±0.348,43.408±0.257 and(8.713±0.106)%,(11.396±0.147)%,(14.110±0.117)%,respectively,the difference were statistically significant between two groups(F=43.436,P<0.001;F=79.751,P<0.001).Conclusion:Silencing AIB1 gene can inhibit breast cancer cell proliferation and promote its apoptosis.

关 键 词：乳腺癌扩增基因1 基因沉默 三阴性乳腺癌 抑制 凋亡 

分 类 号：R737.9[医药卫生—肿瘤]