摘要
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10μM、100μM和1000μM浓度的BH3作用于H1299细胞,72h后,用RT-PCR检测Bcl-2、Capase3和Bcl-xL的表达变化、测定Capase3和Capase9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、Capase3和Bcl-xL的表达表现出相应变化,Capase3和9的活性也有增高的趋势。BH3对细胞有明显的抑制作用,24h后,抑制率分别为2.34%、17.5%、24.46%;48h后,抑制率分别为8.72%、20.33%、45.17%;72h后,抑制率分别为13.11%、38.90%、50.24%。BH3(10μM、100μM和1000μM)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Ca-pase3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 （10μM, 100μM and 1000μM）, 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 facilita ting the expression of caspase 3 to promote apoptosis may be one of anti-tumor mechanisms for BH3 in H1299.
出处
《国际检验医学杂志》
CAS
2011年第5期541-542,584,共3页
International Journal of Laboratory Medicine
