目的 研究肽BH3抗人非小细胞肺癌细胞株H1299的作用,并探讨其作用机制.方法 体外培养H1299细胞,取10 μM、100 μM和1 000 μM浓度的BH3作用于H1299细胞,72 h后,用RT-PCR检测Bcl-2、Capase 3和Bcl-xL的表达变化、测定Capase 3和Capase 9的活性并观察体内外细胞生长的抑制情况.结果 RT-PCR结果 显示,不同浓度的BH3作用后,Bcl-2、Capase 3和Bcl-xL的表达表现出相应变化,Capase 3和9的活性也有增高的趋势.BH3对细胞有明显的抑制作用,24 h后,抑制率分别为2.34%、17.5%、24.46%;48 h后,抑制率分别为8.72%、20.33%、45.17%;72 h后,抑制率分别为13.11%、38.90%、50.24%.BH3(10 μM、100 μM和1 000 μM)对裸鼠移植性非小细胞肺癌细胞株H1299也有抑制作用,3个剂量组的抑瘤率分别为43.85%,54.92% 和59.02%.结论 肽BH3有促进H1299细胞凋亡的作用.降低高表达的Bcl-2和Bcl-xL基因,升高Capase 3基因,可能是BH3抗人非小细胞肺癌的机制之一.%Objective To study the effects of BH3 on human NSCLC cell lines H1299 and its mechanisms. Methods The expression of apoptosis associated genes Bcl-2、Capase 3 and Bcl-xL were observed by RT-PCR.;The direct measurements of caspase 3 and 9 activity were made using colorimetric protease assay kits; MTT assay and weighing the tumor were applied to confirm the inhibition effect. Results Analysis of gene expression showed that BH3 induced caspase-dependent signaling pathways of apoptosis and inhibited Bcl-2 and Bcl-xL gene. Direct measurement of caspase activity showed that the degree of activation of caspase by BH3 was significantly higher when compared with control group. After treatment with BH3 ( 1OμM、 1OOμM and 1OOOμM) , inhibitory effect on the transplanted H1299 was observed. The IR was 43. 85 ％％ , 54. 92％％ and 59. 02 ％ , respectively. Conclusion BH3 can inhibit the growth of human NSCLC cell lines H1299. . Inhibiting the excessive expression of bcl-2 gene .Bcl-xL gene and facilitating the expression of caspase 3 to promote apoptosis may be one of antitumor mechanisms for BH3 in H1299.
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