肿瘤坏死因子相关凋亡诱导配体诱导结肠癌HCT-116细胞凋亡和炎性反应相关基因表达的研究

目的探讨肿瘤坏死因子相关凋亡诱导配体（TRAIL）对结肠癌细胞凋亡和炎性反应相关基因表达的影响。方法HCT-116细胞经10μg／L和100μg／L重组人类TRAIL蛋白处理后,应用荧光实时定量PCR和试剂盒测定凋亡相关基因（Bcl-2、Bad、caspase-3和-8）和炎性反应相关基因（TNF—α,IL-1β,COX-2）的表达水平。结果经10μg几和100μg／L重组人类TRAIL蛋白孵育处理24h后．HCT-116细胞的凋亡率分别为27．4％和45．9％。同时抗凋亡基因Bcl-2、促凋亡基因Bad以及凋亡指示基因caspase-3和caspase-8的表达均显著上调。且100μg／L组的上调幅度均高于10μg／L组（P〈0．05）。TRAIL蛋白处理后,炎性反应相关基因TNF-α,IL-1β,COX-2的表达亦显著上升．且100μg／L处理组的上升幅度均高于10μg／L组（P〈0．05）。结论TRAIL重组蛋白能够诱导结肠癌细胞凋亡以及炎性反应的发生。

Study of tumor necrosis factor-related apoptosis-inducing ligand-induced apoptotic and inflammatory gene expressions in colon cancer cell line

Objective To study the effect of recombinant tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on the expression of apoptosis and inflammatory related genes in human colon cancer cell line HCT-116.Methods After 24-hour treatment with recombinant human TRAIL protein,the expressions of apoptosis-related genes (Bcl-2,Bad,caspase-3,and caspase-8) and inflammation-related genes (TNF-α,IL-1β,and COX-2) were measured by real-time PCR and appropriate kits in HCT-116.Results After treatments of 10 μg/L and 100 μg/L recombinant human TRAIL proteins,the apoptotic rates of HCT-116 cells were 27.4％ and 45.9％,respectively.Expressions of anti-apoptosis gene Bcl-2,pro-apoptosis gene Bad and apoptotic markers caspase-3 and caspase-8were significantly up-regulated,which was more significant in the group of 100 μg/L treatment(P＜0.05).Moreover,after TRAIL treatments,expressions of inflammation-related genes TNF-α,IL-1,COX-2were also dramatically increased,and 100 μg/L treatment group showed higher up-regulation(P＜0.05).Conclusions Recombinant TRAIL protein induces both apoptosis and inflammation of human colon cancer cells.