【摘要】 目的证实苯乙基异硫氰酸酯(phenethyl isothiocyanate,PEITC)诱导人急性髓系白血病(AML)M2白血病细胞系Kasumi-1细胞凋亡的效应,初步探讨PEITC诱导Kasumi-1细胞凋亡的机制。方法培养人M2白血病细胞株Kasumi-1细胞,CCK-8法检测不同浓度PEITC处理后Kasumi-1细胞的增殖抑制率;流式细胞术检测药物诱导细胞凋亡和活性氧(reactive oxygen species,ROS)生成的情况;Real-time PCR及Western blot检测HO-1以及凋亡相关基因caspase3、caspase8、caspase9mRNA及蛋白水平的变化。结果 CCK-8结果显示PEITC作用Kasumi-1细胞24h、48h、72h后,细胞增殖抑制率呈现浓度-时间依赖性。流式细胞术显示PEITC能够诱导Kasumi-1细胞的凋亡。Real-time PCR及Western blot结果显示PEITC处理细胞后,HO-1表达下降,而凋亡相关基因Caspase3、Caspase8、Caspase9的表达上调。DCFH-DA探针法显示PEITC作用Kasumi-1细胞后,ROS生成增多。结论 PEITC能够促进Kasumi-1细胞凋亡,呈时间剂量梯度依赖关系;PEITC诱导Kasumi-1细胞凋亡可能通过的机制包括下调HO-1表达促进ROS生成及凋亡相关基因Caspase3、Caspase8、Caspase9的激活。更多还原

【Abstract】 Objective To investigate the effect of Phenethyl isothiocyanate in inducing acute myelocytic leukemia(AML)Kasumi-1cell line apoptosis,explore the possible mechanisms,thus providing new idea for the treatment of AML.Methods Kasumi-1cells were cultured.The influences of PEITC on the growth inhibition rate of cells were detected by CCK-8.Annexin-V-PI method was used to detect cell apoptosis by flow cytometry.The mRNA and proteins expressions of HO-1and apoptosis-related genes,caspase3,caspase8 and caspase9were detected by Real-time PCR and western blot(WB)respectively.The ROS generation was detected by flow cytometry.Results CCK-8 methods showed that PEITC inhibited the proliferation of Kasumi-1cells in a dose and time-dependent manner.Flow cytometry showed that PEITC could induced Kasumi-1cell apoptosis.Real-time PCR and WB methods showed that the mRNA and protein expressions of apoptosis related genes caspase3,caspase8 and caspase9increased after PEITC treatment.DCFH-DA probe methods showed that PEITC could increased the ROS generation.Conclusion PEITC is found to suppress cell growth and cause apoptosis by down regulating HO-1expression to increase ROS generation and by the activation of caspase3,caspase8 and caspase9.The results of the present study suggest that PEITC may be a potential antitumor compound for AML-M2 therapy.更多还原