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目的:探讨二烯丙基三硫(diallyl trisulfide,DATS)诱导人胃癌SGC-7901细胞凋亡及凋亡过程中c-FLIP的变化及意义。方法采用MTT、western-blot和细胞免疫组化分别检测DATS对SGC-7901细胞的增殖抑制率及c-FLIP的表达情况。光学显微镜观察凋亡形态,流式细胞术检测凋亡率。结果:MTT结果显示,不同浓度DATS(6、8、10、12、14、16mg.L-1)DATS处理SGC-7901细胞24、48小时后,生长抑制率分别为20.4%--79%和36%--90%,DATS抑制作用随浓度及时间逐渐增强(P<0.05)。细胞免疫组化和western-blot显示:9.5mg..L-1DATS处理SGC-7901细胞24、48小时后,与对照组相比,c-FLIP的表达下调(P<0.05)。光学显微镜:通过9.5mg..L-1DATS作用后24、48小时后,胃癌细胞出现了凋亡形态学改变。流式细胞术检测:经过9.5mg..L-1DATS处理SGC-7901细胞24、48小时后,细胞凋亡率逐渐升高。结论:DATS促进SGC-7901细胞凋亡的机制可能与抑制c-FLIP蛋白的表达有关。
Objective: To investigate the changes of c-FLIP in human gastric cancer cell line SGC-7901 induced by diallyl trisulfide (DATS) and its significance. Methods MTT, western-blot and cell immunohistochemistry were used to detect the proliferation inhibition rate and c-FLIP expression of SGC-7901 cells by DATS. The morphological changes of apoptosis were observed by light microscope and the apoptosis rate was detected by flow cytometry. Results: MTT results showed that the growth inhibition rates of SGC-7901 cells treated with different concentrations of DATS (DATS, 6, 8, 10, 12, 14 and 16 mg.L-1) for 24 h and 48 h were 20.4% -79% and 36% - 90%, DATS inhibition gradually increased with concentration and time (P <0.05). Immunocytochemistry and Western-blot showed that the expression of c-FLIP was down-regulated in SGC-7901 cells treated with 9.5 mg · L-1 DATS for 24 and 48 hours (P <0.05). Light microscopy: After passage of 9.5 mg · L-1 DATS for 24 and 48 hours, apoptotic morphological changes occurred in gastric cancer cells. Flow cytometry: SGC-7901 cells treated with 9.5mg. L-1DATS for 24,48 hours, the apoptosis rate gradually increased. Conclusion: The mechanism of DATS promoting SGC-7901 cell apoptosis may be related to inhibiting the expression of c-FLIP protein.