多西紫杉醇对人非小细胞肺癌细胞株A549生长的影响及机制的体外研究

The Inhibitory effect of Docetaxel on the proliferation of human lung cancer cell A549 cells in vitro

目的:探讨多西紫杉醇诱导肺癌细胞株A549细胞的生长抑制作用及其对凋亡相关基因表达的影响。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)法测定整合素α5β1,bcl-2,bax等凋亡相关基因mRNA的表达。结果:(1)多西紫杉醇在1.33～108μg/ml浓度范围内可抑制A549细胞增殖,且此作用与药物浓度、作用时间呈明显相关关系(r=0.947、0.979,P=0.014、0.004)。(2)4～36μg/ml多西紫杉醇作用12～48h,A549细胞可见典型的凋亡形态学改变,且A549细胞凋亡率随药物浓度增加而升高,呈浓度依赖性(r=0.999,P=0.034)。(3)RT-PCR结果显示,多西紫杉醇可明显抑制A549细胞中表达α5整合素亚基基因,使其bcl2/bax比值明显降低,差异均有统计意义(t=10.147、4.583,P=0.01、0.044)。但多西紫杉醇对A549细胞表达VEGF、bF-GF、β1整合素亚基基因无明显影响(P>0.05)。结论:多西紫杉醇可抑制A549细胞增殖,诱导A549细胞凋亡,其机制是通过抑制Bcl-2/Bax表达比例诱导其凋亡,并可能抑制整合素α5β1表达而影响A549细胞的增殖、黏附,以及转移能力。

Objective ： To investigate the inhibition effect of Docetaxd on the proliferation of non-small cell lung cancer A.549 cell and the corresponding mechanism. Methods： MTT assay was used to determine the proliferation inhibition by I）ocetaxel to A549 cell. Annexin- V kit was used to test the early apoptosis of A549 cells induced by Docetaxel. The late apoptotic cells were assessed by propidium iodide （PI） staining. Semi-quantitative RT-PCR was employed to measure the expression of VEGF,bFGF , α5 intergrin submit , β1 intergrin submit ,Bcl-2 and Baxo Results： The proliferation of A549 cells was inhibited significantly by Docetaxel in a concentration-dependent manner and in a time-dependent manner. I）ocetaxe apoptosis of A549 cells in a concentration -dependent manner（ r = 0.999, P = 0.034）. The related level of Bcl-2/Bax and α5 intergrin submit mRNA expression was markedly decreased in A549 cells treated with 12 μg/ml Decetaxel for 24 hours than that in untreated cells（P 〈0.05）,whereas the related expression levels of VEGF,bFGF and β1 intergrin submit mRNA was not significant between two groups （P 〉 0.05）. Conclusions： Docetaxel has the inhibition effect to the proliferation of non-small cell lung cancer A549 cell, and induce apoptosis, the suppressed expression of Bcl-2/Bax may be the mechanisms of apoptosis induction, and the suppressed expression of α5β1 intergrin submit may be influence the proliferation, adhesion and metastasis of non-small cell lung cancer.