SIRT1通过NF-κB通路调节结直肠癌细胞HCT116增殖和活力
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摘要
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-κBcancer cell line HCT116 was used to overexpress SIRT1 by transfection 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 byexpression levels of p-P65and 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 significantly changed after HCT116 treatment with curcumin,an inhibitor of NF-κB(P>0.05).ConclusionSIRT1 reduces the proliferation and viability of colorectal cancer cell line HCT116 by inhibiting the activation of NF-κB.
Objective To investigate whether SIRT1 regulates the proliferation and viability of colorectal cancer cell line HCT116 through NF-κBcancer cell line HCT116 was used to overexpress SIRT1 by transfection 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 byexpression levels of p-P65and 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 significantly changed after HCT116 treatment with curcumin,an inhibitor of NF-κB(P>0.05).ConclusionSIRT1 reduces the proliferation and viability of colorectal cancer cell line HCT116 by inhibiting the activation of NF-κB.
引文
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[17]Suzuki M,Ikeda A,Bartlett JD.Sirt1 overexpression suppresses fluoride-induced p53 acetylation to alleviate fluoride toxicity in ameloblasts responsible for enamel formation[J].Archives of Toxicology,2018,92(3):1283.
[18]Wang C,Chen L,Hou X,et al.Interactions between E2F1 and SirT1 regulate apoptotic response to DNA damage[J].Nature Cell Biology,2006,8(9):1025-1031.
[2]袁瑛,熊斌,徐烨,等.遗传性结直肠癌临床诊治和家系管理中国专家共识[J].实用肿瘤杂志,2018(1):3-16.
[3]刘永斌,保佳玉,葛晔华,等.LPS刺激对人结肠癌SW480细胞基因表达的影响[J].生命科学仪器,2018(3)23-30,37.
[4]李培培,王红英,温丽娜,等.关于NF-κB基因表达及相关信号通路在卵巢癌发生中作用的研究进展[J].中外女性健康研究,2018(2):24-25.
[5]程翔,郭安南.NF-κB抑制促进肿瘤细胞凋亡[J].世界最新医学信息文摘,2018(50):100-101.
[6]王杠杠,宋崴.SIRT1对间充质干细胞分化的影响[J].中国组织工程研究,2018,22(21):3438-3444.
[7]熊伟民,李明峰,宋佳鸿,等.Sirt1促进肝癌细胞增殖和侵袭活性的分子机制[J].现代生物医学进展,2013,13(9):1751-1754.
[8]马威,卢颖,毛俊,等.SIRT1基因在肿瘤中的作用机制[J].国际肿瘤学杂志,2015,42(1):40-42.
[9]Ong ALC,Ramasamy TS.Role of Sirtuin1-p53 regulatory axis in aging,cancer and cellular reprogramming[J].Ageing Research Reviews,2018,43:64-80.
[10]Yang Q,Wang B,Gao W,et al.SIRT1 is downregulated in gastric cancer and leads to G1-phase arrest via NF-κB/Cyclin D1 signaling[J].Molecular Cancer Research Mcr,2013,11(12):1497-1507.
[11]陈丽梅,刘广健,邓艳红,等.结直肠癌肝转移瘤热消融治疗国际专家共识分享[J].中国介入影像与治疗学,2018,15(6):323-326.
[12]齐超.通过生物信息学分析来寻找结直肠癌转移相关的潜在分子机制和关键基因[D].浙江大学,2018.
[13]Damrauer JS,Stadler ME,Acharyya S,et al.Chemotherapy-induced muscle wasting:association with NF-κB and cancer cachexia[J].European Journal of Translational Myology,2018,28(2):7590.
[14]Perkins ND.The diverse and complex roles of NF-κB subunits in cancer[J].Nature Reviews Cancer,2012,12(2):121-32.
[15]Prasad S,Ravindran J,Aggarwal BB.NF-kappa Band cancer:how intimate is this relationship[J].Molecular&Cellular Biochemistry,2010,336(1-2):25-37.
[16]Mitchell SJ,Martinmontalvo A,Mercken EM,et al.The SIRT1 activator SRT1720 extends lifespan and improves health of mice fed a standard diet[J].Cell Reports,2014,6(5):836-843.
[17]Suzuki M,Ikeda A,Bartlett JD.Sirt1 overexpression suppresses fluoride-induced p53 acetylation to alleviate fluoride toxicity in ameloblasts responsible for enamel formation[J].Archives of Toxicology,2018,92(3):1283.
[18]Wang C,Chen L,Hou X,et al.Interactions between E2F1 and SirT1 regulate apoptotic response to DNA damage[J].Nature Cell Biology,2006,8(9):1025-1031.