雷公藤红素抑制A549人肺癌细胞的增殖并增加其凋亡

Celastrol inhibits growth and increases apoptosis of human lung cancer A549 cells

目的探讨雷公藤红素对人A549肺癌细胞增殖的抑制作用及可能机制。方法采用(0、1、2、3、4、5、6)μmol/L雷公藤红素分别处理培养的A549细胞0、24、48、72 h,通过噻唑蓝(MTT)法检测细胞增殖率确定最佳处理浓度和处理时间。后续实验采用(0、1、3)μmol/L雷公藤红素分别处理培养的A549细胞48 h,实时定量PCR检测BAX、Bcl2、胱天蛋白酶3(caspase-3)、caspase-8、caspase-9的mRNA水平;Western blot法检测BAX、Bcl2、裂解型caspase-3 (c-caspase-3)、c-caspase-8、c-caspase-9的蛋白水平。结果雷公藤红素可浓度和时间依赖性地抑制A549细胞生长,(1、3)μmol/L雷公藤红素处理的A549细胞Bcl2的mRNA和蛋白水平降低,BAX、caspase-3、caspase-8、caspase-9的mRNA水平和BAX、c-caspase-3、c-caspase-8、c-caspase-9的蛋白水平增加。结论雷公藤红素可通过线粒体途径增加A549细胞凋亡并抑制其增殖。

Objective To investigate the inhibitory effect of celastrol on the proliferation of human lung cancer A549 cells and its possible mechanism. Methods A549 cells were treated with 0,1,2,3,4,5,6 μmol/L celastrol for 0,24,48,72 hours. The effects of different concentrations and durations on the cell proliferation were evaluated by MTT assay to determine the optimal concentration and treatment time. In the subsequent experiments,A549 cells were treated with( 0,1,3) μmol/L of celastrol for 48 hours. The effects of celastrol on the expression of BAX,B-cell lymphoma 2( Bcl2),caspase-3,caspase-8,caspase-9 mRNA in A549 cells were detected by real-time quantitative PCR. The protein levels of BAX,Bcl-2,cleaved caspase-3( c-caspase-3),c-caspase-8,c-caspase-9 were assessed by Western blot analysis. Results Celastrol inhibited the viability of A549 cells in a concentration-and time-dependent manner. Compared with the group without celastrol treatment,the mRNA and protein level of Bcl2 in A549 cel s treated with( 1,3) μmol/L celastrol decreased significantly,while the expression levels of BAX,caspase-3、caspase-8、caspase-9,c-caspase-3,c-caspase-8,c-caspase-9 increased significantly.Conclusion Celastrol could suppress the proliferation and induced the apoptosis of A549 cel s through mitochondrial pathway.