天然活性分子isatin经p53介导的线粒体途径诱导乳腺癌细胞MCF-7凋亡  

The endogenous oxindole isatin induces apoptosis of MCF-7 breast cancer cells through a p53-mediated mitochondrial pathway

作　　者：宋金莲[1] 马中良[2] 迟晓伟[3] 陈艳萍[1] 侯琳[4] 

机构地区：[1]青岛大学附属妇女儿童医院检验科,山东青岛266034 [2]青岛大学附属医院乳腺外科,山东青岛266000 [3]即墨市妇幼保健院检验科,山东即墨266012 [4]青岛大学医学院生物化学与分子生物学教研室,山东青岛266000

出　　处：《中国药理学通报》2016年第6期773-778,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金项目(No 81472542);青岛市卫生局项目(No 2014WJZD135);青岛市科技局项目(No 13-1-3-67-nsh)

摘　　要：目的探讨吲哚醌(isatin)对乳腺癌细胞MCF-7的抗肿瘤作用及其具体的调控机制。方法不同浓度isatin(0、50、100、200μmol·L^(-1))作用于乳腺癌细胞48 h后,利用细胞荧光染色、RT-PCR、Western blot及流式细胞术等方法检测isatin的促凋亡作用。结果不同浓度isatin处理MCF-7细胞48 h,荧光显微镜观察到细胞出现核染色质聚集、细胞体积缩小及DNA断裂等典型的凋亡形态学改变。RT-PCR和Western blot结果证实,随着isatin浓度的不断增加,p53、Bax mRNA和蛋白表达量也逐渐增加,而Bcl-2 mRNA及蛋白表达下降;流式细胞仪结果提示不同药物浓度作用的细胞膜电位出现不同程度的下降,caspase-9被激活。Western blot结果显示isatin处理组细胞胞质细胞色素C含量明显增加,相反线粒体中细胞色素C含量相应降低。结论 isatin具有明显诱导乳腺癌细胞MCF-7凋亡的作用,其可能的作用机制是经p53的线粒体凋亡通路被激活。Aim To investigate the inducement effect of isatin on apoptosis of breast cancer cell line MCF-7,and explore its detailed mechanism. Methods MCF-7cell lines were exposed to isatin at different concentrations( 0,50,100,200 μmol·L^(-1)) for 48 h. Apoptotic features were demonstrated by nuclei staining with Hoechst 33258. Bcl-2,Bax and p53 mRNA were analyzed by RT-PCR. Caspase-9 activation and mitochondrial depolarization were assayed by flow cytometry.Bcl-2,Bax,p53 and cytochrome c proteins were analyzed by Western blot. Results Isatin induces apoptosis of MCF-7 cells. Furthermore,Bcl-2 expression was decreased and the ratio of Bcl-2 to Bax was significantly decreased by isatin. While,p53 expression relatively decreased. The mitochondrial transmembrane potential was markedly reduced and the release of cytochrome c into the cytosol was increased after treatment with isatin. Simultaneously,caspase-9 was activated.Conclusions Isatin significantly induced the apoptosis of MCF-7 cells in vitro. These results strongly suggest that the p53 dependent mitochondrial pathway is involved in apoptosis.

关 键 词：吲哚醌 乳腺癌细胞 凋亡 p53 Bcl-2 Bax 线粒体膜电位 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R737.9[医药卫生—基础医学]