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目的探索线粒体转录因子A(TFAM)在结肠癌中的表达以及其对结肠癌细胞增殖的影响。方法收集中山大学附属第一医院2013年3月至2013年4月期间的30例结肠癌患者的手术标本,采用real-time PCR技术检测癌组织和癌旁正常结肠组织中TFAM m RNA表达水平。采用real-time PCR和Western blot技术检测TFAM在正常结肠细胞和SW480、HT-29、HCT116三种结肠癌细胞株中的m RNA和蛋白表达水平。探讨上调TFAM表达后对结肠癌细胞株SW480细胞增殖能力的影响。结果 1体内研究结果:结肠癌组织中TFAM m RNA表达比癌旁正常结肠组织中表达明显升高(P<0.000 1)。2体外研究结果:与正常结肠细胞株相比,在SW480、HT-29和HCT116三种结肠癌细胞株中TFAM m RNA表达水平均明显升高(P值分别为0.000 8、0.002 3、0.000 6),其中SW480细胞株增幅最明显。与正常结肠细胞株相比,在SW480、HT-29和HCT116三种结肠癌细胞株中TFAM蛋白表达也均明显升高(P值分别为0.000 2、0.003 8、0.001 6),其中SW480细胞株增幅最明显。用质粒转染上调TFAM基因表达后,MTT实验结果显示,在转染后96 h和120 h时,pc DNA3.1-TFAM-SW480细胞组的增殖率比pc DNA3.1-SW480无义对照组的增殖率明显升高(P<0.000 1)。Brd U实验显示,在转染后48 h时,pc DNA3.1-TFAM-SW480细胞组的增殖率比pc DNA3.1-SW480无义对照组的增殖率明显升高(P<0.001 0)。结论TFAM在结肠癌中表达升高,上调TFAM后,对结肠癌细胞的增殖明显增强,为进一步研究TFAM在结肠癌细胞中的调控作用打下基础。
Objective To explore the expression of mitochondrial transcription factor A (TFAM) in colon cancer and its effect on the proliferation of colon cancer cells. Methods Thirty patients with colon cancer were collected from the First Affiliated Hospital of Sun Yat-sen University from March 2013 to April 2013. Real-time PCR was used to detect the expression of TFAM m RNA in cancer tissue and adjacent normal colorectal tissues. Real-time PCR and Western blot were used to detect the expression of m RNA and protein of TFAM in normal colon cells and SW480, HT-29 and HCT116 colon cancer cell lines. To investigate the effect of TFAM up-regulation on the proliferation of colon cancer cell line SW480. Results1 In vivo results: The expression of TFAM m RNA in colon cancer tissues was significantly higher than that in normal colorectal tissues (P <0.000 1). 2 Results of in vitro studies: Compared with normal colon cell lines, the expression levels of TFAM m RNA in SW480, HT-29 and HCT116 colon cancer cell lines were significantly increased (P values were 0.000 8,0.002 3,0.000 6 ), Which SW480 cell line the most obvious increase. Compared with normal colon cell lines, TFAM protein expression was also significantly increased in SW480, HT-29 and HCT116 colon cancer cell lines (P <0.0002, 0.003 8 and 0.001 6, respectively), of which SW480 cell line The most obvious increase. The results of MTT showed that the proliferation rate of pcDNA3.1-TFAM-SW480 cells was significantly higher than that of pcDNA3.1-SW480 non-sense control groups at 96 h and 120 h after transfection The proliferation rate was significantly higher (P <0.000 1). The BrdU assay showed that the proliferation rate of pcDNA3.1-TFAM-SW480 cells was significantly higher than that of pcDNA3.1-SW480 nonsense control cells (P <0.001 0) at 48 h after transfection. Conclusion The expression of TFAM is increased in colon cancer. The up-regulation of TFAM enhances the proliferation of colon cancer cells, which lays the foundation for the further study on the regulation of TFAM in colon cancer cells.