得力生诱导肺癌细胞株A549细胞凋亡及其对survivin基因表达的影响

目的探讨中成药得力生诱导肺癌细胞株A549细胞凋亡及其对survivin基因表达的影响。方法采用MTT法测定细胞增殖抑制率，在显微镜下观察细胞凋亡的形态特点，应用流式细胞仪检测细胞凋亡率，采用逆转录聚合酶连反应（RT—PCR）法测定survivin mRNA的表达。结果（1）得力生在4mL／L-100mL／L浓度范围内可抑制A549细胞增殖，此作用与药物体积浓度无明显相关关系，但该作用具有时间依赖性（r=0．991，P=0．001）。（2）20mL／L浓度的得力生在体外可诱导A549细胞凋亡，且其作用具有时间依赖性（r=0．997，P=0．049）。（3）RT—PCR结果显示，得力生可明显抑制A549细胞中表达survivin，与对照组相比，差异具有统计学意义（r=4．333，P〈0．05）。结论得力生可抑制A549细胞增殖，诱导A549细胞凋亡，其作用机制可能与抑制survivin基因表达有关。

The Effect of Delisheng in Inducing Apoptosis and Survivin Expression of Human Non-small Cell Lung Cancer A549 Cell in Vitro

Objective To investigate the effect of Delisheng on inducing apoptosis and survivin expression of human non-small cell lung cancer A549 cell. Methods MTT assay was used to determine the proliferation inhibition by Delisheng to A549 cell. Annexin-V kit was used to test the early apoptosis of A549 cells induced by Delisheng. The rate apoptotic cells were assessed by propidium iodide （PI） staining. Semi-quantitative RT-PCR was employed to measure the expression of survivin mRNA. Results （1）4 mL/L-100 mL/L Delisheng significantly inhibited the proliferation of A549 cells in a concentration-independent manner,but in a time-dependent manner（r= 0. 991,P=0. 001）. （2）20 mL/L Delisheng induced apoptosis of A549 cells in vitro,and the function was in a time-dependent manner（r=0. 997,P=0. 049）. （3）RT-PCR proved ：the related level of survivin mRNA expression was markedly decreased in A549 cells treated with 20 mL/L Delisheng for 24 hours than that in untreated cells（t=4. 333,P〈0.05）. Conclusion Delisheng could result in obvious growth inhibitory effects of human non-small cell lung cancer A549 cell and induce apoptosis, the suppressed expression of survivin may be one of the mechanisms of apoptosis induction.