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目的探讨TFⅢB亚单位Bdp1在5-甲基硫代腺苷(methylthioadenosine,MTA)对结肠癌RKO细胞增殖的抑制过程中的作用。方法采用Realtime PCR方法分析TFⅢB亚单位Bdp1基因mRNA水平的变化的转录水平;采用Western Blot方法分析TFⅢB亚单位Bdp1蛋白水平的变化;采用转染及荧光素酶报告基因的检测方法检测Bdp1基因荧光素酶活性。结果 MTA对结肠癌RKO细胞增殖抑制过程中,TFⅢB亚单位Bdp1在mRNA水平(0.441±0.03,P=0.001)和蛋白水平出现了明显的下调,同时报告基因Bdp1基因荧光素酶活性也出现了明显的下调(0.460±0.08,P=0.001)。结论 MTA对结肠癌RKO细胞增殖抑制过程中,TFⅢB亚单位Bdp1蛋白的下调可能起到重要作用。
Objective To investigate the role of TFIIIB subunit Bdp1 in the inhibition of proliferation of RKO cells induced by 5-methylthioadenosine (MTA). Methods Real-time PCR was used to analyze the transcriptional level of Bdp1 mRNA in TFⅢB subunit. Western Blot was used to analyze the level of Bp1 protein in TF Ⅲ B subunit. Transfection and luciferase reporter assay were used to detect the expression of Bdp1 gene fluorescein Enzyme activity. Results MTA inhibited the proliferation of RKO cells by down-regulation of Bdp1 at the level of mRNA (0.441 ± 0.03, P = 0.001) and protein level, and at the same time, the luciferase activity of Bdp1 gene also appeared obvious (0.460 ± 0.08, P = 0.001). Conclusions MTA may play an important role in the down-regulation of B Ⅲp subunit Bdp1 in the process of inhibiting the proliferation of RKO cells in colon cancer.