雷公藤甲素通过调节microRNA21的表达干预K562/A02细胞的化疗敏感性  

Triptolide enhance the sensitivity of doxorubicin via regulating microRNA21 expression in K562/A02 cell line

作　　者：惠璐璐[1] 许文林[1] 沈慧玲[1] 陈巧云[1] 朱小兰[1] 龙璐璐[1] 徐颖[1] 

机构地区：[1]江苏大学附属人民医院中心实验室,江苏镇江212002

出　　处：《江苏大学学报（医学版）》2012年第2期104-109,共6页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(81070423);江苏省医学重点人才资助项目(RC2007034)

摘　　要：目的:研究雷公藤甲素对K562/A02细胞多药耐药性的逆转作用,并探讨其机制。方法:四甲基偶氮唑盐比色(MTT)法检测雷公藤甲素细胞毒性;采用非毒性浓度雷公藤甲素作用于K562/A02细胞,AnnexinV/PI双标法检测细胞凋亡;荧光定量PCR检测microRNA21表达水平;蛋白印迹法检测Bcl-2蛋白表达水平。microRNA21反义寡核苷酸转染K562/A02细胞,观察细胞增长率,凋亡率和Bcl-2表达的变化。结果:非毒性浓度(5 nmol/L)雷公藤甲素明显增强K562/A02细胞对多柔比星的敏感性,并能增强多柔比星诱导细胞凋亡的作用,使K562/A02细胞平均凋亡率由4.3%明显上升到18.5%(P<0.05);雷公藤甲素作用后,microRNA21和Bcl-2蛋白水平降低;microRNA21反义寡核苷酸转染K562/A02细胞后,降低了Bcl-2表达,提高多柔比星敏感性和多柔比星诱导的细胞凋亡。结论:雷公藤甲素增强K562/A02细胞对多柔比星的敏感性,并诱导其凋亡,可能与下调microRNA21水平有关。Objective： To investigate the reversal effects of triptolide on drug resistance in doxorubicin- resistant cells. Methods： Cell viability was measured by MTr assays. The K.562/A02 ceils were treated by triptolid at non-toxicity concentration. The apoptosis of cells were detected by Annexin V/PI methods. The expression of microRNA21 was measured by fluorescence quantitative polymerase chain reaction. Bcl-2 pro- tein level were measured by western blot. After transfected with microRNA21 antisense oligonucleotide, the sensitivity to doxorubicin, the apoptosis of cells and Bcl-2 protein level were detected in K562/A02 cells. Results： Triptolide at non-toxicity concentration significantly enhanced sensitivity of K562/A02 cells to doxorubicin and promoted doxorubicin-induced apoptosis. Levels of microRNA21 and Bcl-2 was significantly decreased after triptolide treatment. Transfection with microRNA21, a significant up-regulation of sensitivity to doxorubicin and a significant down-regulation of Bcl-2 protein was noted in K562/A02 cells. Conclu- sion： Triptolide significantly sensitizes K562/A02 cell to doxorubicin by inducing apoptosis and these effects of triptolide may be due to its down-regulation of microRNA21.

关 键 词：雷公藤甲素 多柔比星 微小RNA21 耐多柔比星人慢性髓系白血病细胞株 B细胞淋巴瘤 白血病-2蛋白 

分 类 号：R733.7[医药卫生—肿瘤]