PS-341与ATO联合用药对K562Bcr-Abl+细胞株细胞周期的影响.docVIP

PS-341与As2O3联合用药对K562Bcr-Abl+细胞株细胞周期的影响 徐瑞达 许啸声 沈子珒 契维 辛秦 指导老师 李稻 上海交通大学医学院03级法文班 病理生理学组 [摘要]本实验研究旨在观察PS-341与As2O3联合用药对Bcr-Abl+慢性粒细胞白血病细胞株K562Bcr-Abl的细胞周期影响。方法：采用细胞培养的方法。在药物作用24h、48h后，收集细胞，进行台盼蓝染色细胞计数、瑞氏染色的细胞形态观察、流式细胞仪检测细胞周期和Annexin-V的凋亡细胞、Western blot的BCR-Abl、P53、NDRG-1蛋白分析。结果：K562Bcr-Abl细胞在PS-341 32nM 与As2O31μM联合用药作用48h后，出现明显的生长抑制，细胞活率明显降低。镜检发现联合用药的K562Bcr-Abl细胞出现胞膜皱缩、空泡和核固缩的细胞增多。细胞周期检测发现G2/M期增多，并出现sub-G1期（为凋亡细胞），Annexin-V、PI染色后检测结果显示凋亡细胞显著增多。Western blot显示联合用药24h时，Bcr-Ablp210表达略有减少；48h时联合用药细胞的Bcr-Ablp210与对照与单独用药细胞比较出现明显减少。结论：为PS-341 32nM与As2O3 1μM联合用药作用48h后，能使K562Bcr-Abl细胞生长抑制、细胞周期阻滞在G2/M期、Bcr-Ablp210表达减少，促进K562Bcr-Abl细胞凋亡。 关键词: K562Bcr-Abl 细胞周期 bcr-abl PS-341 As2O3 Effect of coadministration of PS-341 and As2O3 on the cell cycle of K562Bcr-Abl cell Abstract: The objective of our experiment is to observe the effect of coadministration of PS-341 and As2O3 (arsenic trioxide) on the cell cycle of human chronic myeloid leukemic cell line K562Bcr-Abl. Methods: K562Bcr-Abl cells were cultured and treated with different concentrations of PS-341 and As2O3. After exposed to the agents for 24 and 48 hours, the K562Bcr-Abl cells were collected. Then the growth response was evaluated and the morphologic changes were observed after Wright’s staining. Cell-cycle status and apoptosis (stained by Annexin V–fluorescein isothiocyanate,FITC) were analyzed by flow cytometry. The proteins of Bcr-Abl、P53were determined using a Western blot assay. Results: Coexposure of K562Bcr-Abl cells to PS-341 32nM and As2O3 1μM for 48 hours resulted in a significant inhibition of cell growth and cytotoxicity. The K562Bcr-Abl cells with pyknose increased. The analysis of cell-cycle status showed that the percentage of cells in G2/M phase was enlarged. Moreover, the cells in apoptosic sub-G1 were also appeared. Apoptosis assessment by annexin-V staining demonstrated that there was an increase of the apoptosic K562Bcr-Abl cells. Combined treatment with PS-341 and As2O3 on K562Bcr-Abl cel