毛钩藤碱通过线粒体途径诱导乳腺癌MDA-MB-231细胞凋亡  

Hirsutine induces apoptosis of human breast cancer MDA-MB-231 cells through mitochondrial pathway

作　　者：黄器伟[1] 翟娜娜[1] 黄涛[1] 李道明[2] 

机构地区：[1]黄河科技学院,郑州450099 [2]郑州大学第一附属医院病理科,郑州450052

出　　处：《生理学报》2018年第1期40-46,共7页Acta Physiologica Sinica

基　　金：supported by Science and Technology Development Project of the Science & Technology Department;Henan Province;China(No.162103210003)

摘　　要：本研究旨在观察毛钩藤碱对人乳腺癌细胞的抑制作用,并探讨其分子机制。选取人正常乳腺上皮MCF-10A细胞、乳腺癌MCF-7细胞和MDA-MB-231细胞作为研究对象,采用CCK-8法检测细胞活性,采用流式细胞术检测细胞凋亡和线粒体膜电位(mitochondrial membrane potential,MMP),采用Western blot检测Bcl-2、Bax、cleaved-caspase 9、cleaved-caspase 3以及胞浆中细胞色素C(cytochrome C,Cyt C)的蛋白水平。结果显示,毛钩藤碱可显著降低MCF-7细胞和MDA-MB-231细胞存活率,且该作用呈时间和剂量依赖性(P<0.05);毛钩藤碱作用MCF-7细胞和MDA-MB-231细胞48 h的IC50分别为447.79和179.06μmol/L;毛钩藤碱可诱导MDA-MB-231细胞发生凋亡和MMP去极化(P<0.05),促进线粒体释放Cyt C(P<0.05),激活caspase 9和caspase 3(P<0.05),这些作用均可被线粒体通透性转换孔(mitochondrial permeability transition pore,MPTP)特异性阻断剂环孢素A(cyclosporin A,CsA)显著抑制(P<0.05);此外,毛钩藤碱显著下调MDA-MB-231细胞Bcl-2蛋白水平并上调Bax蛋白水平(P<0.05)。以上结果提示,毛钩藤碱可诱导MDA-MB-231细胞发生凋亡,这可能与其调低Bcl-2/Bax蛋白比值,从而引起MPTP持续开放和Cyt C释放,最终导致caspase 9和caspase 3活化有关。The aim of this study was to investigate the effects of hirsutine on apoptosis of breast cancer cells and its possible mecha- nism. The MCF-10A, MCF-7 and MDA-MB-231 cells were treated with hirsutine at different concentrations for 48 h or incubated with 160 Ixmol/L hirsutine for 24, 48, and 72 h. The MCF-10A cell line is a non-tumorigenic epithelial cell line, and the MCF-7 and MDA-MB-231 are human breast adenocarcinoma cell lines. CCK-8 assay was employed to detect the cell viability. Flow cytometry was used to assay the apoptosis and mitochondrial membrane potential （MMP）. The protein expressions of Bcl-2, Bax, cleaved- caspase 9, cleaved-caspase 3 and cytochrome C （Cyt C） in the MDA-MB-231 cells were detected by Western blotting. The results showed that hirsutine remarkably reduced the viability of MCF-7 and MDA-MB-231 cells in a time- and dose-dependent manner （P 〈 0.05） with IC50 values of 447.79 and 179.06 ktmol/L, respectively. In the MDA-MB-231 cells, hirsutine induced apoptosis and depolarization of MMP （P 〈 0.05）, released Cyt C from mitochondria （P 〈 0.05）, and activated caspase 9 and caspase 3 （P 〈 0.05）. However, these effects induced by hirsutine were all inhibited by cyclosporin A （CsA） （P 〈 0.05）, a specific inhibitor of mitochondrial permeability transition pore （MPTP）. In addition, hirsutine down-regulated the protein level of Bcl-2 and up-regulated the protein level of Bax （P 〈 0.05）. These results suggest that hirsutine may induce apoptosis of human breast cancer MDA-MB-231 cells through decreasing the ratio of Bcl-2 to Bax, opening MPTP, releasing Cyt C from mitochondria, and activating caspase 9 and caspase 3.

关 键 词：毛钩藤碱 乳腺癌 凋亡 环孢素A 线粒体膜电位 

分 类 号：R737.9[医药卫生—肿瘤]