国际医药卫生导报2017，Vol.23Issue(24)：3832-3835,3839,5.DOI:10.3760/cma.j.issn.1007-1245.2017.24.004
miRNA靶向调控CCR7抑制结肠癌细胞增殖及侵袭的研究
Research on microRNA suppressing colon cancer cell proliferation and invasion by targeting CCR7
摘要
Abstract
Objective To investigate the effect of miRNAs on expression of CCR7 in colorectal cancer cells and its influence on colorectal cancer cell proliferation and invasion.Methods The miRNAs targeting CCR7 gene 3'-UTR was predicted by Target Scan,Pictar,and miRanda.5 miRNAs (miR-let7,miR-21,miR-98,miR-145,miR-642) that modulated CCR7 gene expression were selected.3'-UTR of CCR7 gene was amplified through PCR and was inserted in the dual luciferase reporter vector.293 cells were treated with recombinant vector and miRNA by transfection reagent.The dual Luciferase Reporter Assay System was used to evaluate the activity of luciferase.The HCT116 cells treated with miRNAs of migration and invasion were evaluated by wound-heal and transwell assays.The expression of CCR7 was detected by the western blotting.Results 3'-UTR of CCR7 gene was successfully cloned into the psiCHECK vector,which was verified by gel electrophoresis and DNA sequencing methods.3'-UTR of RHD gene may have a binding site of miRNA.Compared with 293 cells treated with the miRNA NC,the luciferase activity of 293 cells cotransfected with recombinant vector and miRNA was decreased.The miR-let7,miR-21 could significantly inhibit the migration and invasion of colorectal cancer cells,and down-regulate the expression of CCR7 in HCT116 cells.Conclusion 3'-UTR of RHD gene dual luciferase combinant vector was successfully constructed.miRNA (miR-let7 and miR-21) could down-regulate expression of CCR7 gene.The expression of CCR7 gene was down-regulated by miR-let7 and miR-21 in colorectal cancer cells,and it may be closely related to the migration and invasion of colorectal cancer.关键词
CCR7基因/结肠癌/miRNA/双荧光素酶报告系统Key words
CCR7 gene/Colorectal cancer/miRNA/Dual luciferase reporter引用本文复制引用
林汉利,陈小欢,王晓玲,曾浩涛..miRNA靶向调控CCR7抑制结肠癌细胞增殖及侵袭的研究[J].国际医药卫生导报,2017,23(24):3832-3835,3839,5.基金项目
深圳市科创委科技创新项目(JCY20140416094330192) （JCY20140416094330192）
深圳市龙岗区科技发展资金项目(YLWS20150513163509153).Innovation Project of Science and Technology Commission of Shenzhen Municipality (JCY20140416094330192) （YLWS20150513163509153）
Longgang District Science and Technology Plan Project (YLWS20150513163509153) （YLWS20150513163509153）
