摘要
目的:研究蟾蜍毒素诱导人胃癌细胞MGC803凋亡及对凋亡因子Livin、caspase-3的影响.方法:应用MTT法检测10、20、40、80、160nmol/L蟾蜍毒素对胃癌细胞MGC803增殖抑制作用;瑞氏-吉姆萨染色法观察细胞形态学的变化;流式细胞术分析周期阻滞和细胞凋亡;Western blot法检测Livin、caspase-3蛋白的表达.结果:(1)蟾蜍毒素抑制MGC803细胞增殖,24、48及72h的IC50分别为105.25nmol/L、47.92nmol/L、16.52nmol/L;(2)10-80nmol/L蟾蜍毒素能诱导胃癌细胞周期阻滞及凋亡,形态学表现为出现凋亡小体;(3)流式细胞仪检测到80nmol/L组亚二倍体凋亡峰(20.83%±2.72%vs1.67%±0.46%,P=0.006)及20nmol/L组细胞周期G2/M期阻滞(37.81%±3.28%vs20.66%±3.09%,P=0.003);(4)Western blot显示蟾蜍毒素诱导细胞凋亡过程中,Livin蛋白表达分别下调到对照组的73.29%±6.23%,60.10%±7.84%和34.70%±5.76%,同时活化caspase-3.结论:蟾蜍毒素诱导胃癌细胞MGC803细胞凋亡可能与下调Livin蛋白、活化caspase-3蛋白有关.
AIM： To explore the mechanisms by which bufa- lin induces cell apoptosis in human gastric can- cer cell line MGC803. METHODS： After MGC803 cells were treated with 10, 20,40, 80, 160 nmol/L of bufalin for different durations, cell growth was measured by MTT assay; cell morphological changes were evaluated by Wright-Giemsa staining and observed under a light microscope; apoptosis was evaluated by flow cytometry; and the expression of livin and caspase-3 was detected by Western blot. RESULTS： Treatment with bufalin inhibited cell growth, and the 50% inhibitory concentrations （ICs0） at 24, 48 and 72 h were 105.25, 47.92 and 16.52 nmol/L, respectively. Treatment with dif- ferent concentrations of bufalin could efficiently induce apoptosis （80 nmol/L, percentage of cells in the sub-G1 peak： 1.67% ± 0.46% vs 20.83% ± 2.72%, P = 0.006） and cell cycle arrest （20 nmol/L, percentage of cells arrested in the G2/M phase： 20.66% ± 3.09% vs 37.81% ± 3.28%, P = 0.003） of MGC803 cells. The expression of livin protein was down-regulated （73.29% ± 6.23%, 60.10% ± 7.84%, 34.70% ± 5.76% vs 100%, all P 〈 0.05） and caspase-3 protein was activated in MGC803 cells treated with 10, 20 and 80 nmol/L bufalin. CONCLUSION： Bufalin induces MGC803 cell apoptosis via mechanisms associated with activation of caspase-3 and inhibition of livin expression.
出处
《世界华人消化杂志》
CAS
北大核心
2011年第2期116-120,共5页
World Chinese Journal of Digestology
基金
辽宁省科技厅博士科研启动基金资助项目
No.20091051~~
