二烯丙基三硫诱导人胃癌细胞凋亡与钙稳态失衡相关

Role of intracellular Ca^(2+) in diallyl trisulfide induction of apoposis in gastric cancer cells

目的探讨二烯丙基三硫(DATS)对人胃癌MGC803细胞的促凋亡作用及钙稳态失衡与该作用的相关性。方法体外培养MGC803细胞,采用MTT法,Tunnel法及流式细胞光度术分析经DATS处理后,对细胞增殖和凋亡的影响以及细胞内游离钙水平的改变。结果DATS处理后,培养的MGC803细胞增殖抑制。4、8、12、16、24mg·L-1的DATS对细胞生长的抑制率分别为0.231±0.037,0.305±0.036,0.455±0.029,0.607±0.058,0.751±0.019。Tunnel检测结果显示,对照组表达阴性,DATS处理组细胞呈阳性表达。流式细胞分析结果显示DATS呈浓度依赖性诱导胃癌MGC803细胞凋亡,16mg·L-1组凋亡率达0.242。细胞内游离钙水平呈浓度依赖性上升,16mg·L-1组荧光强度为对照组的4倍多。用细胞内游离钙特异性阻断剂BAPTAAM预处理细胞后,细胞内游离钙水平不再上升,且能抑制DATS诱导的凋亡。结论DATS能诱导胃癌MGC803细胞凋亡,DATS诱导胃癌MGC803细胞凋亡的作用与钙稳态失衡相关。

Aim To investigate whether DATS induce MGC803 cell apoptosis and the relationship between apoptosis and Ca^2＋ disruption. Methods MGC-803 cell growth inhibition was measured by MTT assay. Tunnel and flow cytometry methods were used to determine the induction of apoptosis and Ca^2＋ homeostasis disruption. Result MTT assay showed that the inhibitory rates on MGC-803 cell growth of different concentrations of DATS 4,8,12, 16 and 24 mg·L^-1 were 0.231± 0. 037,0. 305±0. 036,0. 455±0. 029,0. 607±0. 058,0. 751± 0. 019 respectively. Flow cytometry analysis showed that treating MGC803 cell with DATS significantly increased the percentage of apoptosis ceilsand intracellular Ca^2＋. Treatment of cells with 1,2-bis （2-aminophenoxye-thane） -N, N, N-tetraacetic acid tetrakis acetoxymethyl ester （ BAPTA-AM ）, cellular Ca^2＋ chelator, resulted in abolishment of the elevation of intracellular Ca^2＋ and blockage of DATS induced apoptotic of MGC-803. Conclusoin DATS could induce apoptosis of MGC-803 ceils through the mechanism of Ca^2＋ homeostasis disruption.