沉默Akt减少乳腺癌MCF-7/ADM细胞对阿霉素的耐药性  

Akt knockdown reduces adriamycin resistance in breast cancer MCF-7/ADM cells

作　　者：王玲[1] 蔡岳华[1] 焦荣华[1] 贺百花[1] 阳宁 WANG Ling;CAI Yue-hua;JIAO Rong-hua;HE Bai-hua;YANG Ning(College of Medicine and Technology,Hunan Institute of Environmental Biology and Technology,Hengyang 421001,Hunan,China)

机构地区：[1]湖南环境生物职业技术学院医药技术学院,湖南衡阳421001 [2]南华大学附属第二医院泌尿外科,湖南衡阳421001

出　　处：《广东医学》2019年第13期1841-1845,共5页Guangdong Medical Journal

基　　金：湖南省教育厅基金资助项目(编号:14C0391)

摘　　要：目的探讨沉默Akt对乳腺癌MCF-7细胞阿霉素(ADM)耐药的影响及相关机制。方法培养人乳腺癌细胞ADM敏感株(MCF-7/S)和耐药株(MCF-7/ADM),Western blot检测Akt、磷酸化Akt(p-Akt)水平。利用小干扰RNA(siRNA)沉默MCF-7/ADM细胞中Akt表达,MTT比色法测定肿瘤细胞对ADM敏感性,流式细胞术分析细胞凋亡情况,Western blot检测Bcl-2、Bax和P-糖蛋白(P-gp)表达。结果MCF-7/ADM细胞p-Akt水平明显高于MCF-7/S细胞(P<0.01)。合成的siRNA可有效沉默MCF-7/ADM细胞中Akt表达。Akt siRNA转染MCF-7/ADM细胞增强ADM敏感性,促进细胞凋亡及Bax表达,并减少Bcl-2、P-gp表达(P<0.01)。结论MCF-7/ADM细胞Akt呈活化状态,抑制Akt部分逆转MCF-7/ADM细胞对ADM耐药性,其机制可能与刺激细胞凋亡及下调P-gp表达有关。Objective To investigate the effects of Akt knockdown on adriamycin(ADM)resistance in breast cancer MCF-7/ADM cells and its underlying mechanisms.Methods Western blot was used to determine the levels of Akt and phosphorylated Akt(p-Akt)in human breast cancer MCF-7/S and MCF-7/ADM cells.Small interfering RNA(siRNA)was applied for knockdown of Akt in MCF-7/ADM cells.Subsequently,the sensitivity of tumor cells to ADM was determined by MTT colorimetric technique.Cell apoptosis was analyzed using flow cytometry.The expression of Bcl-2,Bax and P-glycoprotein(P-gp)was measured by Western blot.Results The level of p-Akt was significantly higher in MCF-7/ADM cells than that in MCF-7/S cells(P<0.01).Synthesized siRNA could effectively knockdown Akt in MCF-7/ADM cells.Transfection of MCF-7/ADM cells with Akt siRNA enhanced ADM sensitivity,promoted cell apoptosis and Bax expression,and decreased Bcl-2 and P-gp expression(P<0.01).Conclusion Akt is activated in MCF-7/ADM cells.Inhibition of Akt partially reverses ADM resistance in MCF-7/ADM cells,which is associated with increased apoptosis and decreased P-gp expression.

关 键 词：乳腺癌 AKT 阿霉素 凋亡 P-糖蛋白 

分 类 号：R737.9[医药卫生—肿瘤] R979.1[医药卫生—临床医学]