低毫安电化学疗法诱导人乳腺癌耐药株MCF-7/阿霉素细胞凋亡及逆转多药耐药的研究  

Low milli-ampere electrochemical therapy reverses multidrug resistance and induces apoptosis on breast cancer MCF-7/adriamycin cell line

作　　者：周炳刚[1] 沈义军[2] 魏昌晟[2] 杨涛[2] 张智[2] 余生林[1] 余建军[1] 

机构地区：[1]宁夏自治区人民医院总院微创乳腺外科,银川750002 [2]宁夏医科大学

出　　处：《中华实验外科杂志》2015年第5期1001-1005,共5页Chinese Journal of Experimental Surgery

基　　金：宁夏自然科学基金资助项目（NZ13176）

摘　　要：目的 探讨低毫安电化学疗法（ECT）对人乳腺癌耐药株MCF-7/阿霉素（ADR）细胞诱导凋亡及逆转多药耐药（MDR）的作用机制.方法 电化学处理细胞后继续培养6h和24h,用噻唑蓝（MTT）法、膜联蛋白V（Annexin V）染色、激光共聚焦显微镜观察ECT对肿瘤细胞的生长抑制、凋亡变化;荧光分光光度法检测细胞内ADR的浓度;实时定量聚合酶链反应（Real-time PCR）法、Western blot法检测多药耐药基因（MDR1）、第10号染色体上缺失与张力蛋白同源的磷酸酯酶基因（PTEN）、磷酸化蛋白激酶B（p-Akt）、半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-3基因mRNA和蛋白表达.结果 ECT能明显抑制MCF-7/ADR细胞的生长,实验组凋亡率比对照组明显增加（P＜0.05）;5 C ECT作用后,细胞内的ADR积累增加了4.61倍,实验组PTEN、Caspase-3的相对表达量及蛋白表达随电量的增加明显增高;5 C时P-糖蛋白（P-gp）的相对表达量为0.293 ±0.013、p-Akt蛋白的相对表达量为0.397±0.020,与空白对照组比较,表达明显下降（P＜0.01）.结论 ECT可抑制MCF-7/ADR细胞生长,诱导凋亡,具有逆转多药耐药性,其机制可能主要与ECT作用Akt基因抑制磷脂酰肌醇3-激酶/（PI3K）/Akt信号通路等参与的凋亡途径相关.Objective To explore the mechanism of reversing mutidrug reisistance and inducing apoptosis on human breast cancer MCF-7/adriamycin （ADR） cell line by electrochemical therapy （ECT）.Methods Methyl thiazol tetrazolium （MTT） assay,Annexin V assay and confocal laser scanning microscope were used to measure the inhibitory rate an the change of apoptosis.Fluorospectrophotometry was usd to measure the change of the concertration of ADR in the cells.Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blotting were used to evaluate the mRNA and protein expression levels of multidrug resistance 1 gene （MDR1）,phosphatase and tensin homologue deleted on chromosometen （PTEN）,protein kinase B （Akt） and Caspase-3 in MCF-7/ADR cells.Results ECT could inhibit growth obviously of the MCF-7/ADR cells,and the apoptosis rate of cells was increased obviously in the treated group as compared with that in the control group （P 〈 0.05）.5 C ECT could obviously increase the intracellular concentration of ADR 4.61 times.With the increases in the power of electricity,the expression of PTEN and cleaved Caspase-3 was obviously higher than in the control group,but the protein expression of Permeability glycoprotein （P-gp） （0.293 ± 0.013）,and p-Akt （0.397 ± 0.020） in 5 C ECT group （P 〈 0.01） was reduced gradually with the increases in the power electricity.Conclusion ECT can inhibit the proliferation of MCF-7/ADR cells,induce apoptosis and reverse MDR probably by inhibiting PI3K/Akt signal pathway.

关 键 词：电化学疗法 乳腺癌 多药耐药 蛋白激酶B基因 

分 类 号：R737.9[医药卫生—肿瘤]