二烯丙基二硫诱导人白血病HL-60细胞凋亡及机制

目的　探讨二烯丙基二硫 (DADS)诱导人白血病HL 6 0细胞凋亡的生物学效应及抗肿瘤机制。方法　通过MTT还原法检测DADS对该细胞系生长的影响 ;用电镜、荧光显微镜、流式细胞仪和细胞凋亡原位检测 (TUNEL)研究DADS诱导的细胞凋亡。SP免疫组化法检测细胞内Bcl 2、Bax蛋白的表达。结果　DADS能抑制HL 6 0细胞的生长 ,DADS处理HL 6 0细胞后 ,电镜下细胞呈凋亡特征性改变 :体积变小 ,核浓缩 ,凋亡小体形成 ,流式细胞仪示不同浓度DADS作用于HL 6 0细胞 ,亚G1期细胞明显增多 ,TUNEL测凋亡指数增加。SP免疫组化结果表明 10mg·L-1DADS处理细胞 2 4h后 ,Bax蛋白表达升高 ,Bcl 2蛋白表达下降。结论　DADS有诱导人白血病HL 6 0细胞凋亡的作用 ,其机制与Bcl 2 /Bax比例下降有关。

Apoptosis of human leukemia cell line HL-60 induced by diallyl disulfide and its molecular mechanism

Aim The aim of this work was to study the effect of diallyl disulfide (DADS) on human Leukemia cell line HL 60, and investgate the mechanisms of its antitumor effect. Methods HL 60 cells growth inhibition was measured by MTT assay. Cell apoptosis was inspected by flow cytometry,TUNEL assay and acridine orange fluorescent staining methods. The protein levels of Bcl 2, Bax were determined using immunohistochemical technique. Results MTT assay showed that DADS significantly inhibited the growth of HL 60 cells. After 24 hours of exposure to DADS, Partial cells presented characteristic morphological changes of apoptosis under the electron microscope, including cell shrinkage, nuclear condensation, and formation of apoptotic bodies. Some typical subdiploid peaks before G 0/G 1 phase were observed. Flow cytometry analysis revealed that the apoptosis rates were increased and the TUNEL assay showed the apoptosis index increased in occordance with increase of concentration of DADS. SP immunohistochemistry revealed that the Bax expression was increased while Bcl 2 expressed was decreased. Conclusion Diallyl disulfide could significantly induce apoptosis of human Leukemia cell HL 60, Apoptosis of tumor cells is closely associated with down regulation of the ratio of bcl 2/bax.