雷帕霉素对人淋巴瘤Raji细胞株增殖影响及其机制的探讨  

Growth suppression of human Burkitt lymphoma cell line Raji cells by Rapamycin

作　　者：王炜[1] 高玉环[2] 王彬[2] 刁兰萍[2] 刘海生[2] 武莉丽[2] 

机构地区：[1]保定市第一中心医院血液科,河北保定071000 [2]河北医科大学第四医院暨河北省肿瘤医院血液科,河北石家庄050011

出　　处：《中华肿瘤防治杂志》2011年第23期1843-1846,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：河北省普通高校强势特色学科项目资助〔冀教高(2005)52号〕

摘　　要：目的:探讨雷帕霉素(宜欣可)对人淋巴瘤细胞株Raji细胞体外增殖及细胞周期的影响,并分析其分子作用机制。方法:采用MTT法检测不同浓度(0、1、5、10、20、40、50和100nmol/L)雷帕霉素作用不同时间(24、48和72h)对Raji细胞增殖的影响。用流式细胞仪测定雷帕霉素对Raji细胞周期分布和凋亡的影响。应用蛋白质印迹法检测雷帕霉素对Raji细胞周期蛋白Cyclin D1、Cyclin E、Cyclin A和p27及凋亡蛋白抑制因子Survivin蛋白表达的影响。结果:雷帕霉素浓度>5nmol/L对Raji细胞增殖有明显的抑制作用(P<0.05)。且呈现明显的剂量-效应和时间-效应依赖关系。雷帕霉素处理后,可明显抑制Raji细胞周期发展(P<0.05),随着药物浓度的加大、作用时间的延长,处于G0/G1期细胞逐渐增多,S期和G2/M期细胞则逐渐减少。但Raji细胞没有发生明显的凋亡现象(P>0.05)。50nmol/L雷帕霉素处理后,Raji细胞的Cyclin D1、Cyclin E、p27表达水平没有明显变化(P>0.05),但Cyclin A和Survivin表达水平被明显抑制(P<0.05)。结论:雷帕霉素通过阻滞细胞周期发展抑制Ra-ji细胞增殖,其作用机制可能与下调细胞CyclinA和Survivin表达有关。OBJECTIVE：To investigate the effects of mTOR inhibitor-rapamycin on proliferation of human Burkitt lymphoma cell line Raji cells,and the molecular mechanism was analyzed.METHODS：Effect of rapamycin in different of concentration（0,1,5,10,20,40,50,100 nmol/L） and for different time groups（24,48,72 h） on proliferation of Raji cells was detected by MTT assay.Apoptosis and cell cycle were analyzed by flow cytometry.The expression of Cyclin D1,Cyclin E,Cyclin A,p27,and Survivin protein was examined by western blot technique in the rapamycin-treated and untreated Raji cell.RESULTS：Rapamycin can inhibit the proliferation of Raji cells at concentration greater than 5 nmol/L（P〈0.05）,in a dose and time dependent manner.After treatment with rapamycin,the number of cells in S and G2/M phase decreased gradually（P〈0.05）,but,increased in G0/G1 phase significantly in a dose and time dependent manners（P〈0.05）.However,it could not cause apoptosis in Raji cells evidently（P〈0.05）.The expression of Cyclin D1,p27 and Cyclin E had no obvious change（P〈0.05）,but the expressions of Cyclin A and Survivin protein in Raji cells were inhibited after treated with rapamycin at the concentrate of 50 nmol/L（P〈0.05）.CONCLUSION：Rapamycin inhibits Raji cells proliferation by cell cycle arrest in G0/G1,which associated with down regulate Cyclin A and Survivin.

关 键 词：雷帕霉素 淋巴瘤 RAJI细胞 细胞增殖 细胞周期蛋白 Survivin 

分 类 号：R733.1[医药卫生—肿瘤] R965[医药卫生—临床医学]