雷公藤红素诱导人肝癌SMMC-7721细胞凋亡研究  

Apoptosis of Human Hepatocellular Carcinoma Cell Line SMMC-7721 Induced by The Celastrol

作　　者：张乙川[1] 刘峰[1] 王俊[1] 徐杰[1] 杨茂辉[1] 李劲[1] 

机构地区：[1]攀枝花学院附属医院肝胆胰脾外科,四川攀枝花617000

出　　处：《中国普外基础与临床杂志》2016年第1期48-51,共4页Chinese Journal of Bases and Clinics In General Surgery

摘　　要：目的研究雷公藤红素对人肝癌细胞系SMMC-7721增殖及凋亡的影响,并初步探讨其可能的作用机理。方法体外培养SMMC-7721细胞,细胞接受雷公藤红素处理后,采用CCK-8实验及Annexin V-FITC/PI双染实验观察其对SMMC-7721细胞增殖及凋亡的影响;采用Caspase-3活性检测试剂盒测定Caspase-3活性及Western blot法检测SMMC-7721细胞中NF-κB蛋白的表达;同时用人肝癌细胞系SMMC-7721进行裸鼠成瘤实验,观察雷公藤红素对裸鼠成瘤的影响。结果雷公藤红素可以抑制SMMC-7721细胞增殖并呈浓度和时间依赖性;Annexin V-FITC/PI双染实验显示随着药物浓度的增加,出现凋亡的细胞明显增加;Caspase-3活性测定实验显示细胞接受雷公藤红素处理后,Caspase-3活性明显增强;Western blot实验显示SMMC-7721细胞接受雷公藤红素处理后,细胞中NF-κB蛋白表达下调,并呈现一定的时间依赖性。裸鼠成瘤实验显示,雷公藤红素可以抑制裸鼠种植瘤的生长。结论雷公藤红素能显著抑制SMMC-7721增殖并诱导其凋亡,并可以抑制裸鼠种植瘤的生长。雷公藤红素可能通过激活Caspase-3通路及抑制NF-κB通路诱导SMMC-7721细胞凋亡。Objective To investigate the effects of celastrol on the growth and apoptosis of huamn hepatoma SMMC-7721 cells, and investigate its preliminary action mechansim. Methods SMMC-7721 cells were cultured in vitro, CCK-8 assay and Annexin V-FITC/PI staining method were conducted to investigate the effects of celastrol on the growth and apoptosis ofhuamn hepatoma SMMC-7721 cells after the cells were treated with drugs, and then the Caspase-3 activity and NF-KB protein expression were determined by Caspase-3 activity determination kit and Western blot. Huamn hepatoma SMMC-7721 cells transplantation tumor models in nude mice were established and the effect of celastrol on the growth of transplantation tumor were observed. Results Celastrol could inhibit the SMMC-7721 cells growth in a dose and time dependent manner. Annexin-V/PI staining showed that SMMC-7721 cells were induced to death with the concentration increasing of celastrol. Caspase-3 activity was measured after treatment with celastrol and the results indicated that the activity of caspase-3 was significantly enhanced. Western blot experiments showed that the expression of NF-KB protein decreased in a time-dependent manner after treatment with celastrol. Celastrol could inhibit SMMC-7721 cells transplantation tumor growth in nude mice. Conclusions Celastrol could inhibit the proliferation of human hepatoma SMMC-7721 cells and induces apoptosis, and inhibit SMMC-7721 cell transplantation tumor growth in nude mice. Celastrol induce apoptosis of SMMC-7721 ceils might through activating Caspase-3 pathway and NF-KB pathway.

关 键 词：雷公藤红素 肝癌细胞 CASPASE-3 NF-ΚB蛋白 

分 类 号：R285[医药卫生—中药学]