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
摘要目的 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10 μg/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(Bel-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10 μg/L和100 μg/L重组人类TRAIL蛋白孵育处理24 h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100 μg/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100 μg/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.
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abstractsObjective 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.
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