二烯丙基三硫诱导人胃癌细胞凋亡相关基因的差异表达

The differential expression of apoptosis-associated genes in human gastric cancer MGC803 cells induced by diallyl trisulfide

目的探讨二烯丙基三硫(diallyl trisulfide,DATS)诱导人胃癌MGC803细胞凋亡的分子机制。方法MTT法检测DATS对MGC803细胞的生长抑制作用,荧光显微镜和流式细胞仪观察DATS作用后的细胞凋亡。人细胞凋亡基因芯片检测DATS作用MGC803细胞的凋亡相关基因表达谱,RT-PCR验证凋亡相关基因。结果MTT结果显示,4、8、12、16、24mg.L-1DATS对MGC803细胞生长的抑制率从11%增至78%,呈明显量效关系(P<0.05)。荧光显微镜下,DATS处理后的MGC803细胞核染色质着橘红色并呈固缩状或片段状。流式细胞仪检测显示,8、12、16、24mg.L-1DATS作用24h后,出现典型亚二倍体峰,平均凋亡率分别为11.4%、23.7%、27.4%、31.1%(P<0.05)。人细胞凋亡基因芯片检测结果表明,16mg.L-1的DATS作用4h后出现16个凋亡相关基因表达差异,RT-PCR证实,SODD基因mRNA表达上调、Apaf-1基因mRNA表达下调(P<0.05)。结论DATS诱导人胃癌MGC803细胞凋亡可能与多个基因和信号转导通路作用有关。

Abstract： Aim To investigate the differential expression of apoptosis-associated genes in human gastric cancer MGCS03 cells induced by diallyl trisulfide （ DATS ）. Methods Growth inhibition against MGC803 cells was assayed by MTT assay; The apoptosis induced by DATS was assessed by Flow cytometry and fluorescent microscope. The apoptosis-associated gene expression of MGCS03 cell treated with DATS was determided by Human Apoptosis Gene Array. Apaf-1 and SODD genes were confirmed by RT-PCR. Results DATS had significant growth inhibitory activity against MGC803 cells, inhibition ratio increased from 11% to78% at4, 8, 12, 16 and24 mg.L-1 for72 h （P 〈 0. 05）. The characteristic apoptotic body can be observed in partial cells under the fluorescent microscope. After treated with 8,12, 16 and 24 mg .L^-1 DATS for 24 h, MGC803 cells presented significant subdiploid peaks and apoptosis ratio were 23.7%, 27.4% and 31.1% at average by flow eytometry, respectively（P 〈0. 05）. There were 16 apoptosis-associated genes happened to express variability, in which the mRNA level of Apaf-1 was up-regulated and that of SODD was down-regulated （ P 〈 0. 05 ）. Conclusions DATS-inducing apoptosis in human gastric cancer cell line MGC803 may be relevant with some specific genes and various transduction pathways.