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MicroRNAs(miRNAs)是一类长度约为22 nt的非编码的调控性小RNA,它们在诸多的生命活动中发挥重要作用,如参与调控细胞的增殖、分化、凋亡以及肿瘤的发生发展.MicroRNA-449a/b(miR-449a/b)是脊椎动物中进化保守的miRNA,作为抑癌基因,参与了许多癌症的发生过程,但其在结肠癌中的作用尚不清楚.本文利用实时荧光定量技术研究了miR-449a/b在结肠癌组织中的表达.利用双荧光素酶报告基因检测系统及Western印迹鉴定miR-449a/b的靶基因.应用MTS法和Transwell分别检测miR-449a/b对结肠癌细胞增殖和迁移的影响.检测组蛋白乙酰化酶抑制剂曲古菌素A(trichostatin A,TSA)对结肠癌细胞中miR-449a/b表达的影响.研究结果表明:与正常结肠组织相比,miR-449a/b在结肠癌组织中低表达;miR-449a/b能够结合到FRA-1 mRNA 3’-非翻译区(3’-untranslated region,3’-UTR),从而抑制结肠癌细胞HCT116内源Fra-1的表达;外源转染miR-449a/b明显抑制结肠癌细胞HCT116的增殖和迁移;并且TSA处理能够诱导结肠癌细胞HCT116中miR-449a/b的表达.以上结果提示:miR-449a/b可能通过抑制靶基因Fra-1的表达,进而抑制结肠癌细胞的增殖和迁移.
MicroRNAs (miRNAs) are a class of non-coding small RNAs of about 22 nt in length that play important roles in many biological activities, such as regulation of cell proliferation, differentiation, apoptosis, and tumorigenesis. MicroRNAs -449a / b (miR-449a / b), a evolutionarily conserved miRNA in vertebrates, is involved in many cancers as a tumor suppressor gene, but its role in colon cancer remains unclear.In this paper, real-time fluorescence quantitative The expression of miR-449a / b in colon cancer tissues was studied by using dual luciferase reporter assay system and Western blotting to identify the miR-449a / b target gene.MTS and Transwell were used to detect miR-449a / b On the proliferation and migration of colon cancer cells.And the effect of histone acetylase inhibitor trichostatin A (TSA) on the expression of miR-449a / b in colon cancer cells was studied.The results showed that: Compared with normal colon MiR-449a / b is downregulated in colon cancer tissues compared with the normal tissue; miR-449a / b is able to bind to the 3’-untranslated region (3’-UTR) of FRA-1 mRNA, Expression of endogenous Fra-1 in human colon cancer cell line HCT116; Transfection of miR-449a / b Inhibit the proliferation and migration of colon cancer cell HCT116, and TSA treatment can induce the expression of miR-449a / b in colon cancer cell HCT116.The above results suggest that miR-449a / b may inhibit the expression of target gene Fra-1 Colon cancer cells proliferation and migration.