miR-183-5p靶向调控A1BG促进结肠癌细胞迁移

miR-183-5p Promotes Colon Cancer Cells Migration by Targeting A1BG

目的探讨miR-183-5p在结肠癌中的表达及其作用。方法运用qRT-PCR检测miR-183-5p在211份配对结肠癌样本及不同结肠癌细胞系中的表达;构建miR-183-5p-mimics慢病毒载体,上调结肠癌细胞系SW620和RKO中miR-183-5p表达水平,通过Transwell实验检测miR-183-5p对结肠癌细胞迁移的影响。探讨miR-183-5p的作用靶点α-1-B糖蛋白(A1BG),应用荧光素酶报告系统验证。Western blot及qRT-PCR检测miR-183-5p对A1BG表达的影响。结果qRT-PCR结果显示,与正常黏膜相比,miR-183-5p在结肠癌组织中高表达(P<0.05)。与人正常结肠上皮细胞NCM460相比,miR-183-5p在结肠癌细胞系LoVo、HCT116、Caco-2、HT-29及DLD-1中高表达(P均<0.05)。过表达miR-183-5p可促进结肠癌细胞(SW620、RKO)的迁移能力(P<0.05)。荧光素酶报告基因结果显示A1BG是miR-183-5p的直接作用靶点。过表达miR-183-5p可抑制A1BG在结肠癌细胞系SW620和RKO的表达水平(P<0.05)。结论miR-183-5p可促进结肠癌细胞的迁移,与靶向调控A1BG有关。

Objective This study aimed to investigate miR-183-5p expression pattern in colon cancer patients and cell lines and further study the function of its promote malignant behavior in colon cancer.Methods The expression patterns of miR-183-5p in colon cancer samples and cell lines were confirmed by real-time PCR.The lentiviral expression vector of miR-183-5p has been successfully constructed and investigate its significance for migration in colon cancer cell lines.The target of miR-183-5p was further verified by luciferase reporter assay.The regulation effects of miR-183-5p on A1BG expression in colon cancer cell lines were detected by Western blot and real-time PCR.Results Results of real-time PCR indicated that miR-183-5p was up-regulated in colon cancer patients,up-regulation phenotype were observed in LoVo,HCT116,Caco-2,HT-29 and DLD-1 cell lines.Up-regulation of miR-183-5p in colon cancer cells significantly promoted migration ability(P<0.05).Luciferase reporter assay indicated that A1BG was a direct target of miR-183-5p(P<0.05).Up-regulation of miR-183-5p in colon cancer cells significantly inhibited A1BG expression levels(P<0.05).Conclusion miR-183-5p can promote migration of colon cancer cells by targeting A1BG.