γ-分泌酶抑制剂DAPT通过下调NOTCH1信号通路抑制结肠癌HCT116细胞增殖的实验研究  

Gamma-secretase inhibitor reduces the proliferation of colon cancer HCT116 cells with down-regulation of NOTCH1

作　　者：尹鹏[1] 胡君[1] 储著凌[1] 霍中华[1] 侯乐伟[1] 吕盛[1] 栾荣刚[1] 胡国强[1] 

机构地区：[1]解放军第四五四医院普外烧伤整形外科,南京江苏210002

出　　处：《西部医学》2015年第11期1616-1619,共4页Medical Journal of West China

基　　金：南京军区科研课题(12mb020)

摘　　要：目的探讨γ-分泌酶抑制剂DAPT能否通过下调NOTCH1信号通路抑制结肠癌HCT116细胞增殖,以及对Hes1基因表达的影响。方法使用不同浓度的DAPT(0μM、1μM和10μM)对结肠癌HCT116细胞进行干预。在不同时间点(0h、24h和48h)使用MTT法对HCT116细胞活性进行测定。在干预48h后,使用RT-PCR对在不同浓度(0μM、1μM和10μM)DAPT干预后细胞NOTCH1和Hes1mRNA的表达水平进行测定。使用western blot对在不同浓度(0μM、1μM和10μM)DAPT干预后细胞NOTCH1和Hes1蛋白表达水平进行测定。结果使用不同浓度DAPT(0μM、1μM和10μM)对结肠癌HCT116细胞进行干预后,细胞活性呈时间依赖性和浓度依赖性降低,差异均存在显著统计学意义(P均<0.05)。不同浓度DAPT(0μM、1μM和10μM)干预48h后,结肠癌HCT116细胞NOTCH1和Hes1mRNA和蛋白的表达水平呈浓度依赖性的下降,差异均存在显著统计学意义(P均<0.05)。结论本实验表明γ-分泌酶抑制剂DAPT可以通过下调NOTCH1信号通路,抑制结肠癌HCT116细胞增殖。Objective To explore whether γ-secretase inhibitor, DAPT, would inhibit the proliferation of colon cancer HCTll6 cells by down-regulation of NOTCH1. Methods Three different concentrations （0μM, 1μM and 10μM） of DAPT were used in our study. At different time points （0h, 24h and 48h）, MTT assay was performed to measure the cell viabilities. RT-PCR was used to analyze NOTCH1 and Hes1 mRNA. The protein expression of NOTCH1 and Hes1 were measured by Western blot. Results The cell viabilities of colon cancer HCT116 cells were concentration-dependently and time-dependently inhibited by DAPT. Meanwhile, the mRNA and protein expression of NOTCH1 and Hes1 were al- so concentration-dependently reduced by DAPT in HCT116 cells, after 48 hours of administration. Conclusion Gamma- secretase inhibitor, DAPT, could inhibits the proliferation of colon cancer HCT116 cells and Hes1 expression possibly by down-regulation of NOTCH1.

关 键 词：Γ-分泌酶抑制剂 DAPT HCT116细胞 NOTCH1 HES1 

分 类 号：R965.3[医药卫生—药理学]