Objective To explore the effect of siRNA on Musashi-1 gene expression of proliferation of human colorectal cancer cell line HCT116. Methods Two specific siRNAs of Musashi-1 were cloned into the vector pGenSil-1 and transfected into HCT116 cells by Li-pofectamineTM 2000. The expression levels of Musashi-1 mRNA were detected by PCR in real time. Cell proliferation was determined by MTT,and the changes in cell cycle were evaluated by flow cytometry. Results The siRNA sequences were successfully inserted into the eu-karyotic expression vector pGenSil-1. The sequence specific siRNA efficiently down regulated the Musashi-1 expression at mRNA levels. MTT results showed that the rate of HCT116 cells was decreased markedly and the number of cells at S stage was obviously decreased. Conclusion The synthesized siRNA can effectively decrease the Musashi-1 gene expression and inhibit cell proliferation.%目的 探讨表达载体介导小干扰RNA(siRNA)对人HCT116结肠癌细胞Musashi-1基因表达的抑制作用,揭示其对结肠癌细胞生物学行为的影响.方法 构建靶向Musashi-1的siRNA质粒表达载体,在脂质体介导下转染HCT116细胞,实时定量PCR检测Musashi-1 mRNA表达水平,MTT法检测细胞增殖活性,流式细胞仪检测细胞周期状况.结果 DNA测序证实2条Musashi-1 siRNA表达载体构建成功,与对照组相比均可有效抑制Musashi-1的表达,细胞增殖能力显著降低.结论 Musashi-1的siRNA质粒表达载体可以有效地阻断HCT116结肠癌细胞Musashi-1的mRNA表达从而抑制细胞增殖,促进细胞凋亡.
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