5-氮杂-2'-脱氧胞苷对多药耐药MCF-7/ADR和KBV/200细胞增殖及细胞周期影响作用机制的研究  

The effects of 5-aza-2’- deoxyc ytidine on proliferation and cell c ycle of MCF-7/ADR cells and KBV/200 cells and their mechanism

作　　者：张梦楠[1] 范冬梅[1] 杨铭[1] 胡蕴慧[1] 李双静[1] 姜琳琳[1] 李真真[1] 张砚君[1] 熊冬生[1] 

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,实验血液学国家重点实验室,天津300020

出　　处：《肿瘤》2012年第11期874-879,共6页Tumor

基　　金：国家自然科学基金资助项目(编号:30971291);国家自然科学基金资助项目(编号:30873091)

摘　　要：目的:探讨5-氮杂-2'-脱氧胞苷(5-aza-2'-deoxycytidine,5-Aza-dC)对肿瘤多药耐药细胞生长抑制率、细胞周期的影响及可能的作用机制。方法:采用MTT法检测5-Aza-dC对人乳腺癌耐药细胞株MCF-7/ADR、口腔表皮癌多药耐药细胞KBV/200及其相应的亲本敏感MCF-7和KB细胞的生长抑制率;FCM法检测不同浓度5-Aza-dC(1、5和10μmol/L)作用72h及5-Aza-dC(10μmol/L)作用24、48和72h后,各细胞周期分布的变化;分别采用实时荧光定量-PCR及蛋白质印迹法检测细胞周期相关蛋白cyclin A、cyclin E和p21waf1/cip1mRNA及蛋白的表达情况。结果:5-Aza-dC对耐药细胞株MCF-7/ADR及KBV/200均呈现明显的生长抑制作用,细胞周期被阻滞在G2/M期,并呈时间及剂量依赖性;细胞周期相关蛋白cyclin A、cyclin E和p21waf1/cip1mRNA及蛋白的表达均上调。结论:5-Aza-dC可以通过使MCF-7/ADR及KBV/200细胞的细胞周期被阻滞在G2/M期,从而抑制肿瘤多药耐药细胞的增殖;其作用机制可能与上调细胞周期相关蛋白cyclin A、cyclin E和p21waf1/cip1的表达有关。Objective： To investigate the effect of 5-Aza-dC （5-aza-2’-deoxycytidine） on cell cycle of multidrug resistant cells and its mechanism. Methods： Inhibitory effects of 5-Aza-dC on proliferation of breast cancer MCF-7/ADR cells and oral epidermoid carcinoma KBV/200 cells were detected by MTT method. The changes of cell cycle distribution were examined by flow cytometry when the cells were treated with 5-Aza-dC at different concentrations for 72 h or treated with the same concentration of 5-Aza-dC （10 μmol/L） for different time. The expression levels of cyclin A, cyclin E and p21waf1/cip1 mRNAs and proteins were detected by real-time fluorogenic quantitative-PCR and Western blotting, respectively. Results： The proliferation of MCF-7/ADR and KBV/200 cells was suppressed significantly after 5-Aza-dC treatment. The cell cycle was arrested at G2/M phase in dose- and time-dependent manners. The expression levels of cyclin A, cyclin E and p21waf1/cip1 mRNAs and proteins were elevated. Conclusion： The cell cycle of MCF-7/ADR and KBV/200 cells can be arrested at G2/M phase which consequently results in the inhibition of proliferation of multidrug resistant cells induced by 5-Aza-dC. The molecular mechanisms may be related with up-regulation of expressions of cyclin A, cyclin E and p21 waf1/cip1 mRNAs and proteins.

关 键 词：乳腺肿瘤 口腔肿瘤 脱氧胞苷 细胞抑制剂 细胞周期蛋白类 

分 类 号：R737.9[医药卫生—肿瘤] R737.85[医药卫生—临床医学]