circ-HIPK3通过调控miR-1207-5p促进结直肠癌细胞增殖和转移的作用机制  

Mechanism for circ-HIPK3 promoting proliferation and metastasis of colorectal cancer cell by regulating miR-1207-5p

作　　者：徐宇飞[1] 曾俊杰 邓翰林 XU Yufei;ZENG Junjie;DENG Hanlin(Department of Surgery,the Fifth People's Hospital of Chengdu,Chengdu 611130,Sichuan,China)

机构地区：[1]成都市第五人民医院普外科,四川成都611130

出　　处：《贵州医科大学学报》2022年第7期852-857,共6页Journal of Guizhou Medical University

基　　金：四川省医学科研课题计划(Q18064)。

摘　　要：目的探究环状RNA同源域相互作用蛋白激酶3(circ-HIPK3)通过调控微小RNA-1207-5p(miR-1207-5p)促进结直肠癌(CRC)细胞增殖和转移的作用机制。方法体外培养人CRC细胞系HCT166细胞,分别以pcDNA3.1-NC(空白组)与pcDNA3.1-circ-HIPK3(研究组)转染HCT166细胞48 h,于倒置荧光显微镜下观察转染效率,以不做任何处理的HCT166细胞作为对照组;采用荧光定量PCR检测细胞circ-HIPK3及miR-1207-5p mRNA表达,采用MTT实验及划痕实验分别检测细胞增殖及迁移能力,双荧光素酶实验检测circ-HIPK3与miR-1207-5p的靶向作用,Western blot法检测Wnt/β-catenin通路蛋白表达情况。结果研究组细胞增殖促进率、划痕愈合率、细胞中circ-HIPK3、miR-1207-5p mRNA相对表达水平及Wnt、β-catenin蛋白表达水平均高于对照组和空白组,差异有统计学意义(P<0.05),而对照组与空白组上述各项指标比较差异无统计学意义(P>0.05);双荧光素酶实验结果显示circ-HIPK3上存在miR-1207-5p的结合位点。结论circ-HIPK3通过下调CRC中miR-1207-5p的相对表达,促进细胞增殖和转移,其作用机制可能与激活Wnt/β-catenin通路有关。Objective To explore the mechanism of circular RNA homology domain interacting protein kinase 3(circ-HIPK 3)to promote the proliferation and metastasis of colorectal cancer cells(CRC)via regulating miR-1207-5 p.Methods Human CRC cell line HCT166 cells were cultured in vitro,and then transfected with pcDNA3.1-NC(served as blank group)or pcDNA3.1-circ-HIPK3(served as research group)for 48 h,respectively.HCT166 cells were selected as control group.Fluorescence quantitative PCR(qPCR)was used to detect the expression levels of cellular circ-HIPK3 and miR-1207-5p.MTT assay and scratch assay were used to detect cell proliferation and migration ability,respectively.Dual luciferase assay was applied to detect the targeting effect of circ-HIPK 3 on miR-1207-5 p.Western Blot was performed to detect the protein expression levels of Wnt/β-catenin pathway.Results The cell proliferation rate,scratch healing rate,the relative expression levels of cellular circ-HIPK 3,miR-1207-5 p,Wnt,andβ-catenin in the research group were higher than those in e control and blank groups(P<0.05).There was no significant differences in above indicators between control and blank groups(P>0.05).Dual luciferase assay revealed that there was a binding site between circ-HIPK 3 and miR-1207-5 p.Conclusion circ-HIPK 3 promotes cell proliferation and metastasis via downregulating the relative expression of miR-1207-5 p in CRC,and its mode of action may be related to activation of Wnt/β-catenin pathway.

关 键 词：环状RNA同源域相互作用蛋白激酶3 微小RNA-1207-5p 结直肠癌 增殖 转移 WNT/Β-CATENIN通路 

分 类 号：R735[医药卫生—肿瘤]