miR-204-5p对结直肠癌细胞增殖、侵袭、迁移及上皮间质转化的影响

Effects of miR-204-5p on proliferation, migration, invasion and epithelial-mesenchymal transition in colorectal cancer cells

目的:检测miR-204-5p在人结直肠癌与配对正常肠黏膜组织中的表达差异,观察其对结直肠癌细胞生物学行为的影响。方法:PCR检测23例结直肠癌及配对正常肠黏膜组织中miR-204-5p的表达水平,分析其与临床病理参数的关系。平板克隆实验、划痕实验及Transwell小室侵袭实验评估miR-204-5p对结直肠癌细胞生物学行为的影响。Western blot检测上皮间质转化(epithelia-mesenchymal transition,EMT)相关蛋白的表达。生物信息软件预测并验证miR-204-5p的靶基因。结果:与配对正常肠黏膜组织相比,miR-204-5p在癌组织中低表达(P<0.001),其表达水平与TNM分期(χ~2=9.603,P=0.002)、肿瘤大小(χ~2=7.078,P=0.008)及有无淋巴结转移(χ~2=7.340,P=0.007)相关。过表达miR-204-5p能够降低结直肠癌HT29、LoVo细胞增殖(P<0.000 1;P=0.001)、迁移(P=0.031;P=0.0012)及侵袭能力(P=0.008;P<0.001),并能上调E-cadherin的表达(P<0.000 1;P=0.016)、下调Vimentin的表达(P=0.002;P=0.013)。TargetScan、microRNA.org及miR-TarBase预测显示MAPRE2为miR-204-5p的潜在靶基因,过表达miR-204-5p能下调HT29及LoVo细胞中MAPRE2蛋白表达(P=0.001;P<0.000 1)。结论:miR-204-5p在人结直肠癌中低表达,且能抑制结直肠癌细胞的增殖、侵袭、迁移及EMT。

Objective:To investigate the expression of miR-204-5p in human colorectal cancer tissues and paired normal intestinal mucosal tissue,as well as the effects of miR-204-5p on biological behaviors of colorectal cancer cells.Methods:PCR was used to measure the expression of miR-204-5p in 23 colorectal cancer samples and paired normal intestinal mucosal tissue samples,and its association with clinicopathological parameters was ana.lyzed.Colony formation assay,wound healing assay,and Transwell chamber test were used to evaluate the effects of miR-204-5p on the biological behaviors of colorectal cancer cells.Western blot was used to measure the expression of proteins involved in epithelial-mesenchymal transition(EMT).Bioinformatics software were used to predict the target genes of miR-204-5p.Results:Compared with the paired normal intestinal mucosal tissue samples,the colorectal cancer samples had significantly lower expression of miR-204-5p(P<0.001),which was associated with TNM stage(χ2=9.603,P=0.002),tumor size(χ2=7.078,P=0.008),and the presence or absence of lymph node metastasis(χ2=7.340,P=0.007).For colorectal cancer HT29 and LoVo cells,overpression of miR-204-5p reduced proliferation(P<0.000 1;P=0.001),migration(P=0.031;P=0.001 2),and invasion(P=0.008;P<0.001),upregulated the exprssion of E-cadherin(P<0.000 1;P=0.016),and downregulated the expression of vimentin(P=0.002;P=0.013).TargetScan,microRNA.org,and miRTarBase predictions showed that MAPRE2 was a potential target gene of miR-204-5p,and overexpression of miR-204-5p downregulated the protein expression of MAPRE2 in HT29 and LoVo cells(P=0.001;P<0.000 1).Conclusion:miR-204-5p is lowly expressed in human colorectal cancer and can inhibit the proliferation,migration,invasion and EMT of colorectal cancer cell.