敲低乙酰辅酶A羧化酶1对食管鳞状细胞癌KYSE450细胞迁移的影响及机制  

Effect and mechanism of knockdown acetyl CoA carboxylase 1 on migration of KYSE450 cells

作　　者：刘富磊 刘丹辉 唐家萍 刘玉珍[3,4] 赵宝生[3] LIU Fulei;LIU Danhui;TANG Jiaping;LIU Yuzhen;ZHAO Baosheng(Department of Cardiothoracic Surgery,the First People′s Hospital of Zhengzhou,Zhengzhou 450000,Henan Province,China;Department of Emergency,the People′s Hospital of Hebi,Hebi 458030,Henan Province,China;Department of Thoracic Surgery,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China;Research Center for the Life Science,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China)

机构地区：[1]郑州市第一人民医院心胸外科,河南郑州450000 [2]鹤壁市人民医院急诊科,河南鹤壁458030 [3]新乡医学院第一附属医院胸外科,河南卫辉453100 [4]新乡医学院第一附属医院生命科学研究中心,河南卫辉453100

出　　处：《新乡医学院学报》2024年第5期407-411,共5页Journal of Xinxiang Medical University

基　　金：河南省医学科技攻关计划省部共建重点项目(编号:SBGJ202102188)。

摘　　要：目的探讨敲低乙酰辅酶A羧化酶1(ACC1)对食管鳞状细胞癌KYSE450细胞迁移能力的影响及机制。方法将对数生长期食管鳞状细胞癌KYSE450细胞随机分为shNC组、shACC1组、shNC+AEB071组及shACC1+AEB071组,shNC组KYSE450细胞转染空白质粒载体,shACC1组KYSE450细胞转染慢病毒质粒载体,shNC+AEB071组KYSE450细胞转染空白质粒载体后加入5μL浓度为2 mmoL·L^(-1)的蛋白激酶C抑制剂AEB071(终浓度为5μmoL·L^(-1)),shACC1+AEB071组KYSE450细胞转染慢病毒质粒载体后加入5μL浓度为2 mmoL·L^(-1)的蛋白激酶C抑制剂AEB071(终浓度为5μmoL·L^(-1))。Transwell法检测各组KYSE450细胞迁移能力;显微镜下观察各组KYSE450细胞形态学变化;Western blot检测4组KYSE450细胞中乙酰辅酶A羧化酶1(ACC1)、组蛋白H3(H3)、乙酰化的组蛋白H3第9赖氨酸(H3K9Ac)及上皮-间质转化(EMT)标志物β-连环蛋白(β-catenin)、波形蛋白(Vimentin)以及锌指转录因子(Snail)等的表达。结果shACC1组KYSE450细胞迁移数显著高于shNC组(P<0.05);shNC+AEB071组与shNC组KYSE450细胞迁移数比较差异无统计学意义(P>0.05);shACC1+AEB071组KYSE450细胞迁移数显著低于shACC1组(P<0.05)。shNC组KYSE450细胞呈椭圆形上皮样细胞形态,shACC1组KYSE450细胞呈类似于梭形的间质样细胞形态,shNC+AEB071组和shACC1+AEB071组KYSE450细胞呈椭圆形上皮样细胞形态。shACC1组KYSE450细胞中ACC1相对表达量显著低于shNC组(P<0.05),β-catenin、Vimentin、Snail的相对表达量及H3K9Ac/H3比值显著高于shNC组(P<0.05);shNC+AEB071组与shNC组KYSE450细胞中ACC1、β-catenin、Vimentin、Snail相对表达量及H3K9Ac/H3比值比较差异无统计学意义(P>0.05);shACC1+AEB071组KYSE450细胞中β-catenin、Vimentin、Snail的相对表达量及H3K9Ac/H3比值显著低于shACC1组(P<0.05);shACC1+AEB071组与shACC1组KYSE450细胞中ACC1相对表达量比较差异无统计学意义(P>0.05)。结论敲低ACC1可促进食管鳞状细胞癌KYSE450细胞的迁移,从而Objective To explore the effect and the possible mechanism of knockdown acetyl CoA carboxylase 1(ACC1)on the migration of KYSE450 cells.Methods KYSE450 cells during the logarithmic phase were randomly divided into the shNC group,shACC1 group,shNC+AEB071 group,and shACC1+AEB071 group.The KYSE450 cells in the shNC group were transfected with empty plasmid;the KYSE450 cells in the shACC1 group were transfected with lentiviral plasmid;the KYSE450 cells in the shNC+AEB071 group were transfected with empty plasmid and then added with 5μL of 2 mmoL·L^(-1)protein kinase C(PKC)inhibitor AEB071(final concentration 5μmoL·L^(-1));The KYSE450 cells in the shACC1+AEB071group were transfected with lentiviral plasmid and then added with 5μL of 2 mmoL·L^(-1)PKC inhibitor AEB071(final concentration 5μmoL·L^(-1)).The migration of KYSE450 cells was detected by Transwell assay.The morphological changes of KYSE450 cells were observed under the microscope.The expression levels of ACC1,histone H3(H3),histone H3 lysine 9 acetylation(H3K9Ac),and epithelial-mesenchymal transition(EMT)markers such asβ-catenin,Vimentin and Snail were measured by Western blot.Results The migration of KYSE450 cells in the shACC1 group was significantly higher than that in the shNC group(P<0.05);there was no significant difference in the migration ability of KYSE450 cells between the shNC+AEB071 group and the shNC group(P>0.05);the migration of KYSE450 cells in the shACC1+AEB071 group was significantly lower than that in the shACC1 group(P<0.05).The KYSE450 cells in the shNC group revealed an elliptical epithelial-like cell morphology;the KYSE450 cells in the shACC1 group exhibited a spindle-like interstitialcell morphology;the KYSE450 cells in the shNC+AEB071 and shACC1+AEB071 groups showed an elliptical epithelial-like cell morphology.The relative expression level of ACC1 in KYSE450 cells in the shACC1 group was significantly lower than that in the shNC group(P<0.05),while the relative expression levels ofβ-catenin,Vimentin and Snail as well as the

关 键 词：乙酰辅酶A羧化酶1 蛋白激酶C 上皮-间质转化 细胞迁移 

分 类 号：R735.1[医药卫生—肿瘤]