阿托伐他汀钙下调肌微管素相关蛋白3诱导自噬抑制结肠癌HCT116细胞增殖迁移  

Downregulation of MTMR3 by atorvastatin calcium suppresses autophagy and inhibits proliferation and migration of colorectal cancer HCT116 cells

作　　者：刘龙飞[1,3] 刘艳丽 陈燕华 王莉 谢荣俊[1] 莫中成[2] 张森 LIU Long-fei;LIU Yan-li;CHEN Yan-hua(Department of Gastrointestinal Surgery,Nanhua Hospital Affiliated to University of South China,Hengyang(421000),China)

机构地区：[1]南华大学附属南华医院胃肠外科,衡阳421000 [2]桂林医学院,桂林541000 [3]广西医科大学附属第一医院结直肠肛门外科,南宁530000 [4]湘南学院附属医院普外科,郴州423000

出　　处：《中国中西医结合外科杂志》2024年第6期910-918,共9页Chinese Journal of Surgery of Integrated Traditional and Western Medicine

基　　金：湖南省自然科学基金(2020JJ4552)。

摘　　要：目的:探讨阿托伐他汀钙(ATO)诱导自噬抑制结直肠癌(CRC)HCT116细胞增殖与迁移的机制。方法:采用免疫组化检测结直肠癌组织及癌旁组织中肌微管素相关蛋白3(MTMR3)及P62的表达,用0、12.5、25、50、100μmol/L浓度ATO处理结直肠癌HCT116细胞系24 h,采用MTT法检测HCT116细胞活力,使用12.5、25、37.5μmol/L ATO处理HCT116细胞后,进行克隆集落形成实验及细胞划痕实验,并用Western blot法检测其MTMR3及自噬相关蛋白LC3和P62蛋白的表达。ATO联合si-MTMR3、ATO联合自噬抑制剂氯喹及单独氯喹处理结肠癌细胞HCT116,检测HCT116细胞活力、集落形成及迁移能力,并使用Western blot法检测MTMR3、LC3和P62蛋白表达。结果:MTMR3和P62在CRC组织中高表达,经ATO处理的HCT116细胞,药物浓度越高,细胞活力、集落形成和迁移能力越低,其MTMR3及P62蛋白表达亦明显下降,自噬相关蛋白LC3II/I比值逐渐升高,自噬抑制剂氯喹可逆转ATO对HCT116细胞的抑制作用,si-MTMR3与ATO对HCT116细胞有协同抑制作用,并且si-MTMR3可诱导自噬的发生。结论:ATO能抑制结肠癌HCT116细胞的增殖及迁移,其机制可能与通过抑制MTMR3的表达、诱导细胞自噬有关。Objective To investigate the mechanisms underlying the induction of autophagy inhibition and the subsequent suppression of proliferation and migration in colorectal cancer(CRC)HCT116 cells by atorvastatin calcium(ATO).Methods Firstly,immunohistochemical staining was performed to detect the expression of MTMR3 and P62 in CRC tissues and adjacent non-cancerous tissues.Secondly,HCT116 cells were treated with ATO at concentrations of 0,12.5,25,50,and 100μmol/L for 24 hours,and cell viability was assessed using the MTT assay.Thirdly,HCT116 cells were treated with ATO at concentrations of 12.5,25,and 37.5μmol/L,and colony formation and cell migration assays were conducted.Western blot analysis was performed to examine the expression of MTMR3,as well as autophagy-related proteins LC3 and P62.Lastly,HCT116 cells were treated with ATO in combination with si-MTMR3 or autophagy inhibitor chloroquine,or with chloroquine alone.Cell viability,colony formation,and migration capacity of HCT116 cells were assessed,and the expression of MTMR3,LC3,and P62 proteins was examined using Western blot analysis.Results The results showed that MTMR3 and P62 were highly expressed in CRC tissues.Treatment of HCT116 cells with ATO resulted in decreased cell viability,colony formation,and migration capacity,which correlated with reduced expression of MTMR3 and P62,as well as an increase in the LC3II/I ratio,indicative of autophagy induction.Moreover,the inhibitory effects of ATO on HCT116 cells were reversed by the autophagy inhibitor chloroquine.Additionally,si-MTMR3 exhibited a synergistic inhibitory effect with ATO on HCT116 cells and induced autophagy.Conclusion ATO suppresses proliferation and migration of HCT116 cells in CRC,potentially through the downregulation of MTMR3 expression and induction of cellular autophagy.

关 键 词：阿托伐他汀钙 肌微管素相关蛋白3 自噬 结直肠癌 

分 类 号：R735[医药卫生—肿瘤]