摘要
目的探讨微小RNA-152(miR-152)靶向血管紧张素Ⅱ1型受体(AGTR1)对肝癌HCCLM3细胞上皮间质转化(EMT)及肾素-血管紧张素系统(RAS)的影响。方法将培养的HCCLM3细胞分为未转染组(未处理)、阴性对照组(转染阴性对照序列)和miR-152组(转染miR-152模拟物)。采用实时荧光定量PCR检测miR-152和血管紧张素转换酶(ACE)、血管紧张素Ⅱ(AngⅡ)、AGTR1 mRNA表达,Transwell^(TM)小室检测细胞侵袭和迁移,Western blot法检测细胞波形蛋白(vimentin)、神经钙黏蛋白(N-cadherin)、上皮钙黏蛋白(E-cadherin)和AGTR1蛋白表达,双荧光素酶报告基因实验检测miR-152和AGTR1的靶向关系。结果与未转染组或阴性对照组相比,miR-152组细胞miR-152表达水平和E-cadherin蛋白表达水平明显升高,而ACE、AngⅡ、AGTR1 mRNA表达水平和侵袭细胞数、迁移细胞数以及vimentin、N-cadherin、AGTR1蛋白表达水平均明显降低;双荧光素酶报告基因实验检测结果证实miR-152可与AGTR1靶向结合。结论miR-152可能通过靶向下调AGTR1表达抑制肝癌HCCLM3细胞的EMT及RAS。
Objective To investigate the effects of microRNA-152(miR-152)targeting at angiotensinⅡtype 1 receptor(AGTR1)on the epithelial mesenchymal transition(EMT)and renin-angiotensin system(RAS)of HCCLM3 human hepatocellular carcinoma cells.Methods The cultured HCCLM3 cells were divided into untransfected group(untreated),negative control group(transfection negative control sequence)and miR-152 group(transfected miR-152 mimic).The expressions of miR-152,angiotensin converting enzyme(ACE),angiotensinⅡ(AngⅡ)and angiotensinⅡtype 1 receptor(AGTR1)mRNAs were detected by real-time fluorescence quantitative PCR.Cell invasion and migration were detected by Transwell^(TM) assay.The expression of vimentin,N-cadherin,E-cadherin and AGTR1 were tested by western blot.The targeting relationship between miR-152 and AGTR1 were examined by double luciferase reporter assay.Results Compared with the untransfected group or the negative control group,the expression levels of miR-152 and E-cadherin protein in the miR-152 group significantly increased,while the expression levels of ACE,AngⅡ,AGTR1 mRNA,the number of invaded cells,the number of migrating cells,and the protein expression levels of vimentin,N-cadherin,and AGTR1 decreased significantly.The results of double luciferase reporter gene assay confirmed that miR-152 can target binding with AGTR1.Conclusion miR-152 may inhibit EMT and RAS of HCCLM3 cells by targeting down-regulation of AGTR1 expression.
作者
全裔
杨峻
秦韬
王秀娟
胡玉芳
QUAN Yi;YANG Jun;QIN Tao;WANG Xiujuan;HU Yufang(Laboratory of Affiliated Hospital,Affiliated Hospital of Guilin Medical College,Guilin 541001,China;School of Medical laboratory,Affiliated Hospital of Guilin Medical College,Guilin 541001,China;Department of Radiology,Affiliated Hospital of Guilin Medical College,Guilin 541001,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2022年第9期819-824,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
2019年度广西高校中青年教师科研基础能力提升项目(2019KY0536)。
