Rev-erb激动剂SR9009通过降低自噬抑制人结肠癌细胞系HCT116增殖  

Rev-erb agonist SR9009 inhibits the proliferation of human colon cancer cell line HCT116 through reducing autophagy

作　　者：石光 宁娜 侯晓鸿 孙聪 原媛 王丽[1] 王晓晖[1] SHI Guang;NING Na;HOU Xiao-hong;SUN Cong;YUAN Yuan;WANG Li;WANG Xiao-hui(Department of Pathology,School of Basic Medicine,Shanxi Medical University,Taiyuan 030001,China;Laboratory of Morphology, School of Basic Medicine, Shanxi Medical University, Taiyuan 030001, China)

机构地区：[1]山西医科大学基础医学院病理教研室,山西太原030001 [2]山西医科大学基础医学院形态学实验室,山西太原030001

出　　处：《基础医学与临床》2020年第11期1494-1499,共6页Basic and Clinical Medicine

摘　　要：目的探讨血红素调节核受体(Rev-erb)激动剂SR9009对结肠癌细胞系HCT116增殖活性的作用及其机制。方法培养HCT116细胞,向培养液中加入Rev-erb激动剂SR9009,倒置相差显微镜观察细胞的增殖;选择SR9009的适宜浓度与作用时间,采用CCK-8法检测细胞活性;Real-time PCR检测Rev-erbα、Rev-erbβ以及自噬基因Beclin1、LC3、p62的转录;Western blot检测Rev-erbα、Rev-erbβ、Beclin1、LC3和p62的蛋白表达。结果1)与对照组相比,SR9009显著抑制HCT116细胞的增殖(P<0.05),SR9009的有效作用浓度为20μmol/L,作用时间为24 h。2)SR9009显著提高HCT116细胞Rev-erbα和Rev-erbβ的转录(P<0.01)以及蛋白表达水平(P<0.05)。3)SR9009显著降低HCT116细胞Beclin1和LC3的表达水平(P<0.05),引起自噬底物p62累积。4)在SR9009处理基础上,用3-甲基腺嘌呤(3-MA)抑制自噬,HCT116细胞活性降低(P<0.05);用雷帕霉素提高自噬,HCT116细胞活性回升(P<0.05)。结论Rev-erb激动剂SR9009可以通过降低自噬来抑制结肠癌细胞系HCT116的增殖活性。Objective To investigate the effect of hemeregulated nuclear receptor(Rev-erb)agonist SR9009 on the proliferation and potential mechanism of human colon cancer cell line HCT116.Methods HCT116 cells were cultured and treated with Rev-erb agonist SR9009 with proper dosagee and treatment time.The cell proliferation was observed with a phase contrast microscopve.The cell viability was tested using a CCK-8 kit.The mRNA transcription of Rev-erbα,Rev-erbβand autophagy genes Beclin1,LC3 and p62 were detected by real-time PCR.The protein expression of Rev-erbα,Rev-erbβ,Beclin1,LC3 and p62 were detected by Western blot.Results1)Rev-erb agonist SR9009 inhibited the HCT116 cell proliferation as compared with control group(P<0.05),and the right dose of SR9009 was 20μmol/L and the treatment time was 24 hours.2)SR9009 significantly increased Rev-erbαand Rev-erbβmRNA transcription(P<0.01)and protein expression(P<0.05)in HCT116 cells.3)R9009 significantly reduced the expression of autophagy genes Beclin1 and LC3 in HCT116 cells(P<0.05),causing autophagy protein p62 accumulation.4)HCT116 cell viability was further decreased with an autophagyinhibitor 3-methyladenine(3-MA)after SR9009 treatment(P<0.05),the cell viability was restored by an autophagy activator rapamycin(P<0.05).Conclusions Rev-erb agonist SR9009 inhibits proliferation and cell viability of colon cancer cell line HCT116 by reducing autophagy.

关 键 词：REV-ERB SR9009 结肠癌 自噬 

分 类 号：R73-36[医药卫生—肿瘤] R735.3[医药卫生—临床医学]