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目的研究5-甲基硫代腺苷(MTA)对结肠癌Hct116细胞增殖的影响及其相关机制。方法采用MTT比色法及软琼脂克隆形成试验分析MTA对结肠癌Hct116细胞增殖的影响,采用Realtime PCR的方法分析RNA聚合酶Ⅲ依赖基因(tRNAs、5S rRNA)及TFⅢB亚单位TBP、Bdp1和Brf1基因mRNA水平的变化,并采用Western Blot的方法分析了TFⅢB亚单位Bdp1蛋白水平的变化。结果 MTA对结肠癌Hct116细胞具有明显的增殖抑制作用,在这一过程中,RNA聚合酶Ⅲ依赖基因(tRNAs、5S rRNA)的mRNA水平明显下调;同时TFⅢB亚单位Bdp1在mRNA水平和蛋白水平也出现了明显下调。结论 MTA对结肠癌Hct116细胞具有明显增殖抑制作用;RNA聚合酶Ⅲ依赖基因转录的下调、TFⅢB亚单位Bdp1蛋白的下调可能在这一抗肿瘤过程中起到重要作用。
Objective To investigate the effects of 5-methylthioadenosine (MTA) on the proliferation of colon cancer Hct116 cells and its related mechanisms. Methods MTT assay and soft agar colony formation assay were used to analyze the effect of MTA on the proliferation of Hct116 cells. Realtime PCR was used to analyze the expression of RNA polymerase Ⅲ-dependent genes (tRNAs, 5S rRNA) and TFⅢB subunits TBP, Bdp1 and Brf1 The changes of Bclp1 protein level in TFⅢB subunit were analyzed by Western Blot. Results MTA had a significant inhibitory effect on colon cancer Hct116 cells. In this process, mRNA level of RNA polymerase Ⅲ dependent genes (tRNAs, 5S rRNA) was significantly down regulated. At the same time, mRNA and protein level of TF Ⅲ B subunit Bdp1 There has been a significant downward adjustment. Conclusions MTA can inhibit the proliferation of Hct116 cells. The down-regulation of RNA polymerase Ⅲ-dependent gene transcription and the down-regulation of B Ⅲp subunit Bdp1 protein may play an important role in this anti-tumor process.