摘要
近年来的研究显示泛素 蛋白酶体通路在细胞凋亡调控中发挥了重要的作用。本实验就蛋白酶体抑制剂Z LLL CHO与经典的凋亡诱导剂足叶乙甙 (VP16)对白血病细胞系M 0 7e及TF 1的联合效应进行了初步研究。应用MTT法及台盼蓝拒染法显示Z LLL CHO及VP16联合作用于M 0 7e及TF 1细胞 ,可增强两种药物单独的细胞杀伤效应。流式细胞术检测提示单独应用Z LLL CHO及VP16均可使S +G2 /M期细胞比例增加 ,两者联合应用导致细胞凋亡峰比例的显著增高。通过对M 0 7e细胞裂解物做免疫印迹检测 ,发现在Z LLL CHO及VP16单独作用中均导致Bcl 2蛋白被酶解为 2 2kD的片段 ,而联合作用时Bcl 2的酶解现象具有叠加效应。结论 :Z LLL CHO与VP16联合作用较单独作用有更强的细胞杀伤及诱导细胞凋亡活性 ,细胞周期S +G2 /M期比例的增加及Bcl 2酶解片段的增加是导致蛋白酶体抑制剂Z LLL CHO及VP16对白血病细胞系联合效应的可能机制。
Recent researches indicate that ubiquitin-prot ea some pathway plays an important role in apoptosis regulation. Proteasome inhibit ors induce apoptosis in many kinds of neoplastic cells, thus provide a great opp ortunity for exploring synergy of proteasome inhibitors and other apoptosis-ind ucing agents. In this study, the effect of the proteasome inhibitor Z-LLL-CHO combined with etoposide (VP16) on leukemic cell lines M-07e and TF-1 was inves tigated by MTT assay, trypan blue exclusion, flow cytometry and Western blot. Th e results showed that the combination of Z-LLL-CHO and VP16 was much more effe ctive than either agents alone in promoting cytotoxicity in both cell lines eval uated. Accumulation of cells in S+G 2/M phase of the cell cycle was observed in the cells treated with VP16 and Z-LLL-CHO alone, while apparent increase of s ub-G 0/G 1 fraction was detected in cells treated with combination of the agents. The cleavage of Bcl-2 into a shortened 22 kD fragment was detected in M -07e cells e xposed to either agents alone, and the fraction of 22 kD fragment was increased in the cells treated with conbination of the agents. In conclusion, the combinat i on of Z-LLL-CHO and VP16 enhanced their individual cytotoxic effect by induci ng apoptosis, in which increase of S+G 2/M fraction in cell cycle as well as th e enhanced cleavage of Bcl-2 are the possible mechanism of the additive effect on leukemic cells by Z-LLL-CHO and VP16.
出处
《中国实验血液学杂志》
CAS
CSCD
2003年第5期485-489,共5页
Journal of Experimental Hematology
