Rheb过表达在白血病细胞株HL-60和K562中的作用  

Effects of Rheb Overexpression in HL-60 and K562 leukemia cell lines

作　　者：徐乔竹 汪晓敏 王放放[2] 高亚男 张英驰 鞠振宇[3] 程涛 袁卫平 刘汉芝 

机构地区：[1]中国医学科学院、北京协和医学院血液病医院、血液学研究所,天津300020 [2]天津医科大学,天津300071 [3]浙江杭州师范大学衰老研究所,浙江杭州310036

出　　处：《中国实验血液学杂志》2013年第2期268-272,共5页Journal of Experimental Hematology

基　　金：科技部973项目(编号2011CB964801,2012CB966600,2010CB945204);国家自然科学基金(编号81090414,81130074,81070390);天津市科委自然基金(编号10JCZDJC19500,11JCZDJC27900)

摘　　要：mTOR信号通路是细胞内重要的生命活动枢纽,它通过抑制细胞凋亡,促进细胞增殖来调控细胞生命活动。Rheb可以激活mTOR信号通路,进而参与多种肿瘤的发生发展。本研究以髓系白血病细胞株为研究对象,探讨Rheb在HL-60和K562中的作用及相关机制。本研究利用逆转录病毒技术使髓系白血病细胞株HL-60和K562过表达Rheb;利用Western blot和Real-Time PCR分别检测HL-60和K562细胞中Rheb的蛋白表达水平及mRNA表达水平;利用CCK-8法检测细胞活力;利用Annexin V-PE和7-AAD双染检测细胞凋亡。结果表明:成功构建了Rheb过表达的HL-60和K562细胞株,并发现Rheb过表达可以促进细胞生长;进一步研究发现,Rheb过表达加快细胞进入G2/M期(P<0.01),但不影响细胞凋亡。结论:Rheb通过加快细胞周期进程促进HL-60和K562细胞增殖。mTOR （ mammalian target of rapamycin） is the center for cellular activities. It controls many cell activi- ties via inhibiting apoptosis and promoting cell growth. Rheb can activate mTOR signaling pathway and participate in genesis and development of multiple cancers. This study was purposed to explore the underlying role of Rheb in human myeloid leukemia by using the myeloid leukemia cell lines. Two myeloid leukemia cell lines HL-60 and K562 overex- pressing Rheb were established with retrovirus containing Rheb. The mRNA and protein expressions of Rheb were deter- mined by Real-Time PCR and Western blot respectively. Cell proliferation rate was examined by CCK-8 assay and apop- tosis rate was analyzed using Annexin V and 7-AAD double-staining. The results showed that Rheb was overexpressed in both HL-60 and K562 cell lines. The Rheb overexpression cell lines were successfully established. It is found that over- expression of Rheb could promote cell growth. Furthermore, the overexpression of Rheb could accelerate cells entering into G2/M phase （P 〈0.01 ）, while did not affect the apoptosis. It is concluded that Rheb overexpression promotes my- eloid leukemia cell proliferation through accelerating cell cycle progression.

关 键 词：RHEB mTORC信号通路 AML HL-60 K562 

分 类 号：R733.7[医药卫生—肿瘤]