Objective To observe inhibitory effect of mesaconitine in WenYang herbs on leukemia K562 cells and K562 resistant daunorubicin cell line(K562/DNR)and possible molecular mechanism.Methods Effect of different concentrations of mesaconitine on proliferation and apoptosis of K562 and K562/DNR cells was detected by using MTT and flow cytometry.After treatment for 72 h,effect of different concentrations of mesaconitine on C/EBPα,Caspase-3 and p53 in K562 and K562/DNR cells was detected by real-time PCR.Results The proliferation of K562 and K562/DNR cells was significantly inhibited by mesaconitine in concentration-and time-dependent manner(P＜0.05).Mesaconitine significantly induced apoptosis of K562 and K562/DNR cells,and the apoptosis rate was increased with increasing of mesaconitine concentration(P＜0.05).Real-time PCR results showed that after treatment with 50 μmol/L mesaconitine for 72 h,C/EBPα was significantly decreased,Caspase-3 and p53 were significantly increased in K562 and K562/DNR cells as compared with the normal control group(P＜0.05).Conclusion Mesaconitine can induce apoptosis of K562 cells and K562/DNR cells,and the underlying mechanism is related with downregulation of C/EBPα and upregulation of Caspase-3 and p53.%目的 观察温阳药新乌头碱对白血病细胞K562和K562耐柔红霉素细胞株(K562/DNR)的抑制作用及可能的分子机制.方法 应用MTT法,结合流式细胞术检测不同浓度新乌头碱对K562和K562/DNR增殖和凋亡的影响.通过Real-time PCR检测不同浓度新乌头碱作用72 h对K562和K562/DNR细胞C/EBPα、Caspase-3、p53凋亡相关基因表达的影响.结果 K562和K562/DNR细胞的增殖抑制率随着新乌头碱浓度增加及作用时间延长而上升,呈浓度、时间依赖性(均P＜0.05).新乌头碱能引起K562和K562/DNR细胞凋亡,且随着药物浓度增加,凋亡率也逐渐升高,差异均有统计学意义(均P＜0.05).Real-time PCR结果显示,经50μmol/L新乌头碱作用72 h,K562和K562/DNR细胞的C/EBPα表达下调,Caspase-3、p53表达上调,与正常对照组相比差异均有统计学意义(均P＜0.05).结论 新乌头碱能够引起K562和K562/DNR细胞发生凋亡,并且凋亡机制可能与下调C/EBPa基因及上调Caspase-3、p53基因有关.
展开▼
