低毫安电化学疗法抑制磷酸肌醇3激酶／蛋白激酶B信号通道抗人乳腺癌细胞株MDA—MB231侵袭迁移的机制  

The electrochemotherpy inhibited migration and invasion of breast cancer - MDA - MB231 cell line through phosphatidylinositol 3 kinase/protein kinase B signal pathway

作　　者：周炳刚[1] 魏昌晟[2] 张智[3] 张颂[3] 苏振民[3] 沈义军[3] 王健[1] 

机构地区：[1]宁夏自治区人民医院微创乳腺外科,银川750002 [2]甘肃省肿瘤医院乳腺外科,兰州730050 [3]宁夏医科大学研究生院,银川750004

出　　处：《中华实验外科杂志》2016年第4期917-921,共5页Chinese Journal of Experimental Surgery

基　　金：宁夏自然科学基金（NZ13176）

摘　　要：目的探讨电化学疗法（ECT）对人乳腺癌MDA—MB231细胞株抗侵袭迁移的作用机制。方法采用噻唑蓝（MTT）法检测1～15C的ECT和（0．05—1．00）μmol／LMK2206作用MDA—MB231细胞24h后的生长抑制率；Transwell实验检测5CECT对细胞侵袭、迁移能力改变；用反转录一聚合酶链反应（RT．PCR）、Westernblot法检测1～10CECT作用MDA-MB231细胞24h后血管内皮生长因子（VEGF）、基质金属蛋白酶（MMP）-2、第10号染色体上缺失与张力蛋白同源的磷酸酯酶基因（PTEN）、蛋白激酶B（Akt）基因mRNA和蛋白的表达。结果Transwell实验证实5CECT作用后MDA．MB231细胞侵袭、迁移能力下降；RT—PCR结果显示5C、10C的ECT组与空白对照组比较，随着ECT电量的提高，侵袭基因VEGF、MMP-2的表达量逐渐降低，5CECT组VEGF基因的相对表达量为0．591±0．038，MMP-2基因为0．601±0．045，与空白对照组比较差异有统计学意义（P〈0．01）；磷酸肌醇3激酶（P13K）／Akt信号通路中的PTEN基因表达量逐渐增高而Akt基因的表达量逐渐降低；相同抑制率的ECT组（1C）和MK2206组（0．05μmol／L）比较，两组降低VEGF、MMP-2、Akt基因表达量的差异无统计学意义（P〉0．05），MK,2206组能增高PTEN基因的表达量。Westernblot检测结果显示3C、5C、10C不同库仑剂量的ECT治疗组与空白对照组比较，随着ECT库仑剂量的提高VEGF、MMP-2蛋白表达量逐渐降低[3CECT组VEGF的相对表达量为0．426±0．072，MMP-2的相对表达量为0．214±0．016，与空白对照组比较差异有统计学意义（P〈0．01）]，P13K／Akt信号通路中的磷酸化Akt（P—Akt）蛋白表达量逐渐降低，PTEN蛋白表达量逐渐增高，总Akt基因表达量无明显变化；相同抑制率的ECT组1C和MK2206组0．05±mol／L比较，ECT组更能降低VEGF、MMP-2的蛋白表达量且更能增高PTEN的蛋白表达量，但MK2206组更能降低p-Akt的蛋白表达量，对总Akt蛋Objective To explore the mechanism of electrochemical therapy （ECT） through phos- phatidylinositol 3 kinase（ PI3K）/protein kinase B （Akt） signal pathway inhibiting migration and invasion of breast cancer cell lines. Methods MTr assay was used to measure the inhibtion rate of 1 - 15 C ECT coulomb and （0. 05 - 1.00） μmol/L MK2206 on breast cancer MDA - MB231 cell lines after 24 h. Tran- swell migration and matrigel invasion assays were performed to evaluate the ability of 5 C ECT migration and matrigel invasion respectively, reverse transcriptase -polymerase chain reaction （RT- PCR） and Western blotting were used to evaluate the mRNA and protein expression levels of vascular endothelial growth factor （VEGF） , matrix metalloproteinase （MMP） -2, phosphatase and tensin homologue deleted on chromo- someten （PTEN）, Akt and phosphorylated Akt （p - Akt） in MDA - MB231 cells. Methods Transwll showed ECT could inhibit the ability of cell migration and invasion obviously. RT - PCR showed the rela- tive expression levels of VEGF and MMP -2 in 5 C and 10 C ECT groups were reduced gradually （for VEGF： 0. 591 ±0. 038, and for MMP -2： 0. 601 ±0. 045 in 5 C ECT group; for VEGF, 0. 453 -±0. 003, and for MMP -2： 0. 410 ±0. 064 in 10 C ECT group,P 〈0. 01 ）. The relative gene expression of Akt was reduced gradually, but PTEN increased gradually. The expression levels of VEGF, MMP - 2 and Akt in the ceils treated separately by 1 C ECT and 0. 05 μmol/L MK2206 showed no significant difference at the same inhibition rate. Western blotting showed expression levels of VEGF and MMP - 2 in the 5 C, 5 C and 10 C ECT groups were reduced gradually （for VEGF, 0. 426 ±0. 046 and for MMP -2, 0. 223 ±0. 018 in 5 C ECT group,P 〈0. 01 ）. The protein expression of p -Akt was also reduced gradually, but PTEN in- creased. The total Akt protein expression was unchanged （ P 〉 0. 05 ）. At the same inhibitory rate of 1 C ECT and MK2206 （0. 05 μmoL/L）, the ECT group was more effective th

关 键 词：电化学疗法 乳腺癌 侵袭性 磷酸肌醇3激酶/蛋白激酶B信号通路 

分 类 号：R285.5[医药卫生—中药学]