阿西替尼对结肠癌HCT-116细胞增殖、凋亡及自噬的影响研究  

Influence of Acetinib on proliferation,apoptosis and autophagy of colon cancer HCT-116 cells

作　　者：潘晟[1] 梅文超 黄林飞[1] 夏甘霖 陶艳娥 徐竞[1] 李俊[1] PAN Sheng;MEI Wenchao;HUANG Linfei;XIA Ganlin;TAO Yan'e;XU Jing;LI Jun(Department of Gastrointestinal Surgery,Puren Hospital Affiliated to Wuhan University of Science and Technology,Wuhan 430081,China)

机构地区：[1]武汉科技大学附属普仁医院胃肠外科,湖北武汉430081

出　　处：《中国肿瘤外科杂志》2020年第5期407-411,共5页Chinese Journal of Surgical Oncology

基　　金：湖北省卫生健康委员会联合基金项目(WJ2019H225)。

摘　　要：目的研究阿西替尼(AXI)对结肠癌HCT-116细胞增殖、凋亡及自噬的影响。方法体外培养结肠癌HCT-116细胞,分为AXI 1组(15.0μmol/L)、AXI 2组(30.0μmol/L)、AXI 3组(60.0μmol/L)及无任何添加的正常对照组(NC组)。四甲基偶氮唑蓝法(MTT)检测各组结肠癌HCT-116细胞增殖情况;AnnexinV-FITC/PI法检测各组结肠癌HCT-116细胞凋亡情况;Western Blot检测结肠癌HCT-116细胞自噬相关蛋白LC3Ⅱ、beclin1,雷帕霉素靶蛋白(mTOR)信号通路中p-mTOR、p-P70S6K蛋白水平、增殖相关蛋白CyclinD1、c-Myc,以及凋亡相关cleaved-caspase3、B细胞淋巴瘤-2(Bcl-2)、Bax蛋白表达水平。结果与NC组比较,AXI 1组和AXI 2组结肠癌HCT-116细胞增殖抑制率、凋亡率、LC3Ⅱ、beclin1、Bax、cleaved-Caspase3蛋白表达水平依次增加(P<0.05),p-mTOR、p-P70S6K、Cyclin D1、c-Myc、Bcl-2蛋白表达水平依次减少(P<0.05)。AXI 2组和AXI 3组结肠癌HCT-116细胞增殖抑制率、凋亡率、LC3Ⅱ、beclin1、p-mTOR、p-P70S6K、CyclinD1、c-Myc、cleaved-caspase3、Bcl-2、Bax蛋白表达水平比较,差异无统计学意义(P>0.05)。结论AXI可通过抑制mTOR通路激活,诱导结肠癌HCT-116细胞自噬,抑制其增殖,促进其凋亡,初步揭示了AXI对结肠癌细胞的作用机制,为结肠癌的靶向治疗提供了新思路。Objective To explore the effects of Acetanil(AXI)on the proliferation,apoptosis and autophagy of human colon cancer HCT-116 cells.Methods Colon cancer HCT-116 cells were cultured in vitro and divided into 15.0μmol/L(A group),30.0μmol/L(B group)and 60.0μsmol/L(C group).The proliferation of HCT-116 cells was detected by MTT assay.Annexin V-FITC/PI was used to detect the apoptosis of HCT-116 cells in colon cancer.And Western blot was used to detect the expression of autophagy-related proteins,including LC3Ⅱ,beclin1,p-mTOR,p-P70S6K protein levels in the target protein(mTOR)signaling pathway of rapamycin,proliferation-related proteins CyclinD1,c-Myc,apoptosis-related cleaved-caspase3,B-cell lymphoma-2(Bcl-2)and Bax in colon cancer cells.Results Compared with NC group,the inhibition rate,apoptotic rate of HCT-116 cells,LC3Ⅱ,beclin1,Bax,cleaved-Caspase3 protein expression levels of colon cancer HCT-116 cells in group A and group B increased by degress(P<0.05).The expression levels of p-mTOR,p-P70S6K,Cyclin D1,c-Myc and Bcl-2 protein decreased by degress(P<0.05).There was no significant difference in the expressions of proliferation,apoptotic rate of colon cancer HCT-116 cells,LC3Ⅱ,beclin1,p-mTOR,p-P70S6K,CyclinD1,c-Myc,cleaved-caspase 3,Bcl-2,Bax protein between group B and group C(P>0.05).Conclusions AXI induces the autophagy of colon cancer HCT-116 cellsby inhibiting the activation of mTOR pathway,which preliminarily reveals the mechanism of action of AXI on colon cancer cells and provides new ideas and theoretical references for the treatment of colon cancer.

关 键 词：结肠癌 阿西替尼 HCT-116细胞 增殖 自噬 凋亡 

分 类 号：R735.35[医药卫生—肿瘤]