SIRT1通过NF-κB通路调节结直肠癌细胞HCT116增殖和活力

SIRT1 regulates proliferation and viability of colorectal cancer cell line HCT116 via NF-κB pathway

目的探讨SIRT1是否通过NF-κB通路来调节结直肠癌细胞HCT116点增殖和活力。方法选用结直肠癌细胞HCT116,通过转染SIRT1质粒来过表达SIRT1,此为过表达组;通过siRNA来敲低SIRT1,此为敲低组;用MTT法、CCK-8法和克隆形成试验来测定HCT116细胞的增殖和活力情况;通过免疫印迹试验来检测NF-κB通路相关蛋白的表达情况。结果过表达组中,p-P65、p-IKKalpha的表达量明显降低(P<0.05),而P65和IKKalpha的总体表达量没有明显差异(P>0.05),HCT116的生长速度、增殖速率以及细胞活力明显降低(P<0.05);敲低组中,p-P65、p-IKKalpha的表达量明显上升(P<0.05),而P65和IKKalpha的总体表达量没有明显差异(P>0.05),HCT116的生长速度、增殖速率以及细胞活力明显上升(P<0.05)。在过表达SIRT1的同时,用NF-κB的抑制剂Curcumin处理HCT116后,NF-κB通路相关蛋白的表达量以及HCT116的生长速度和增殖速率无明显变化(P>0.05)。结论 SIRT1通过抑制NF-κB的激活来降低结直肠癌细胞HCT116增殖和活力。

Objective To investigate whether SIRT1 regulates the proliferation and viability of colorectal cancer cell line HCT116 through NF?κB pathway.Methods Colorectal cancer cell line HCT116 was used to overexpress SIRT1 by trans?fection of SIRT1 plasmid,which is an overexpression group;SIRT1 was knocked down by siRNA,which is a knockdown group;MTT assay,CCK?8 assay and colony formation assay to determine the proliferation and viability of HCT116 cells;the expression of NF?κB pathway?associated proteins was detected by immunoblotting.Results The expression levels of p?P65 and p?IKKalpha were significantly decreased in the overexpression group(P<0.05),but there was no significant difference in the total expression of P65 and IKKalpha(P>0.05).The growth rate and proliferation rate of HCT116 and The cell viability was significantly decreased(P<0.05).The expression of p?P65 and p?IKKalpha was significantly increased in the knockdown group(P<0.05),but there was no significant difference in the total expression of P65 and IKKalpha(P>0.05).The growth rate,proliferation rate and cell viability of HCT116 increased significantly(P<0.05).At the same time of over?expressing SIRT1,the expression of NF?κB pathway?related protein and the growth rate and proliferation rate of HCT116 were not signifi?cantly changed after HCT116 treatment with curcumin,an inhibitor of NF?κB(P>0.05).Conclusion SIRT1 reduces the pro?liferation and viability of colorectal cancer cell line HCT116 by inhibiting the activation of NF?κB.