miR-223在结直肠癌细胞中对RhoB调控作用的研究  

Study on the regulatory effect of miR-223 on RhoB in colorectal cancer cells

作　　者：杨鹏春 白卫哲 杨静芳 汪菁 晏彩华 戴迟兵[1] YANG Pengchun;BAI Weizhe;YANG Jingfang;WANG Jing;YAN Caihua;DAI Chibing(Department of Digestive Internal Medicine,Affiliated Renhe Hospital of Three Gorges University,Yichang,Hubei 443001,China)

机构地区：[1]三峡大学附属仁和医院消化内科

出　　处：《重庆医学》2020年第5期689-694,700,共7页Chongqing medicine

基　　金：三峡大学科研基金项目（KJ 2013A019）

摘　　要：目的探讨miR-223在结直肠癌细胞中对RhoB的调控作用。方法采用PITA、Targetscan等软件在线预测miR-223的靶基因;采用实时荧光定量聚合酶链反应(qRT-PCR)检测miR-223在人结直肠癌细胞株(SW480、HCT-116)中的表达;采用双荧光素酶报告系统证实RhoB是miR-223的靶基因;通过细胞转染,分析miR-223在人结直肠癌细胞株中对RhoB的影响。结果miR-223的表达量在SW480中显著低于HCT-116(P<0.05);PmiR-RhoB-wt和miR-223 mimics共转染组与PmiR-RhoB-wt和NC共转染组比较,前者的荧光素酶活性明显下降(P<0.05),PmiR-RhoB-mut和miR-223 mimics共转染组与PmiR-RhoB-mut和NC共转染组比较,两组间的荧光素酶活性差异无统计学意义(P>0.05);将miR-223 mimics或miR-223 mimics-NC转染至SW480细胞,miR-223 inhibitor或miR-223 inhibitor-NC转染至HCT-116细胞,发现RhoB mRNA表达水平差异均无统计学意义(P>0.05),但是RhoB蛋白的表达差异有统计学意义(P<0.05)。结论miR-223可能靶向作用于RhoB影响结直肠癌的发展及转移。Objective To investigate the regulation effect of miR223 on RhoB in colorectal cancer cells.Methods The target genes of miR-223 were predicted online by PITA and Targetscan software;the expression of miR-223 in human colorectal cancer cell lines(SW480,HCT-116)was detected by qRT-PCR;the dual luciferase reporter system was used to confirm that RhoB is the target gene of miR-223;the effect of miR-223 on RhoB in human colorectal cancer cell lines was analyzed by the cell transfection.Results The expression level of miR-223 in SW480 was significantly lower than in HCT-116 by qRT-PCR(P<0.05);Compared with the PmiR-RhoB-wt and NC co-transfection groups,the PmiR-RhoB-wt and miR-223 mimics co-transfection groups showed a significant decrease in luciferase activity(P<0.05),but there was no statistically significant difference in luciferase activity between PmiR-RhoB-mut and miR-223 mimics co-transfection group and PmiR-RhoB-mut and NC co-transfection group(P>0.05);miR-223 mimics or miR-223 mimics-NC was transfected into SW480 cells,and miR-223 inhibitor or miR-223 inhibitor-NC was transfected into HCT-116 cells,the results showed that there was no statistically significant difference in the RhoB mRNA expression level(P>0.05),but the expression level of RhoB protein had statistically significantly difference(P<0.05).Conclusion miR-223 may targets to act on RhoB,and affect the development and metastasis of CRC.

关 键 词：miR-223 结直肠肿瘤 RHOB 

分 类 号：R735.35[医药卫生—肿瘤]