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

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

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。

Objective To study the effect of recombinant tumor necrosis factor-related apoptosis-indueing 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-8 were 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-1β, COX-2 were 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.