白藜芦醇抑制白血病K562细胞增殖机制的探讨  

Mechanisms underlying resveratrol-induced inhibition of K562 cell proliferation

作　　者：张海燕[1] 都镇先[2] 孟欣[3] 邓娓娓[1] 王华芹[3] 

机构地区：[1]中国医科大学附属第一医院老年病干诊科,辽宁沈阳110001 [2]中国医科大学附属第一医院内分泌科,辽宁沈阳110001 [3]中国医科大学基础医学院生化与分子生物教研室,辽宁沈阳110001

出　　处：《中华肿瘤防治杂志》2012年第19期1460-1463,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：辽宁省科技攻关项目(2010225032);辽宁省教育厅资助项目(2009A765;L2010616)

摘　　要：目的:探讨白藜芦醇(RES)抑制白血病K562细胞增殖的机制及非折叠蛋白应答(UPR)对其机制的影响。方法:选取不同浓度的RES干预K562细胞,并且用RES(100μmol/L)干预K562细胞不同时段;利用MTT法检测细胞存活率;利用流式细胞仪分析细胞周期并检测细胞凋亡率;利用蛋白质印迹法检测凋亡早期蛋白及UPR靶点蛋白表达水平;利用实时定量PCR法检测GRP78和CHOP mRNA的表达。结果:RES导致K562细胞细胞存活率明显减少,并呈剂量依赖方式;RES的生长抑制作用主要是阻断细胞周期中的G1期,G2/M期的变化较小;RES处理的K562细胞没有显著的细胞凋亡。RES可提高GRP78mRNA的表达水平,呈剂量及时效依赖方式;RES对CHOP mRNA水平也有诱导作用,但只有在RES浓度较大时(50μmol/L)或作用时间较长时(8h)才开始上调,并且上调幅度较小。蛋白质印迹分析证实RES提高UPR靶点(GRP78、GRP94、磷酸化eIF2a和剪接作用XBP-1)的蛋白表达水平。结论:RES优先诱导UPR存活分支,表明UPR参与RES抑制白血病K562细胞增殖的机制。OBJECTIVE：To investigate whether unfolded protein response （UPR） involved in mechanisms underly- ing resveratrol-indueed inhibition of K562 cell proliferation. METHODS： K562 cells were treated with different concentra- tion of resveratrol,and 100 μmol/L of resveratrol were selected for treating K562 cells with different period;3-（4,5-dime- thylthiahiazo-2-thiazolyl）-2,5 diphenyl tetrazolium bromide（MTT） assay was used for cell viability assays; Analysis of the cell cycle or detection of eel1 death was confirmed by flow cytometry;marked protein of early apoptosis phase and UPR targets were detected by western blot analysis; the expression of GRP78 and CHOP mRNA were measured by real-time RT PCR. RESULTS： Exposure of K562 leukemia cells to RES resulted in a marked reduction of cell viability in a dose-de- pendent manner;the growth inhibitory effect of RES was the result of a block of cell cycle primarily at G1 phase,to a less- er extent at G2/M phase. Remarkable apoptosis in RES-treated K562 cells was not observed. RES-induced GRP78 ex- pression in a dose-dependent manner;After longer period（8 h） or larger concentrations（50 μmol/L） of RES exposure, CHOP mRNA levels was also upregulated, but with rather minor degree. Several targets of UPR including GRP78, GRP94,phosphorylated elF2a and splicing of XBP-1 were also demonstrated to be increased by western blot analysis. CONCLUSION： UPR involved in mechanisms underlying resveratrol-indueed inhibition of K562 cell proliferation.

关 键 词：K562细胞 半胱氨酸天冬氨酸蛋白酶 代谢 细胞凋亡 药物作用 

分 类 号：R285.5[医药卫生—中药学]