摘要
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 investigate the expression of mitochondrial transcription factor A （TFAM） in colon cancer and the effect of its expression on proliferation of colon cancer cell. Methods Thirty cases of colon cancer in the First Affiliated Hospital of Sun Yat-sen University from March 2013 to April 2013 were studied. TFAM mRNA was detected both in colon cancer tissue and para-cancer tissue by real-time PCR. TFAM mRNA and protein were detected in normal colon cell strain and colon cancer strains SW480, HT-29, and HCT116 by real-time PCR and Western blot, respectively. The proliferation of SW480 cells was evaluated after up-regulating TFAM. Results The expression of TFAM mRNA in the colon cancer tissue was significantly higher than that in the para-cancer tissue （P〈0.000 1）. The expressions of TFAM mRNA were obviously increased in the SW480, HT-29, and HCTll6 cells as compared with the normal colon cell strain （P value was 0.000 8, 0.002 3, and 0.000 6, respectively）, among which the most notable increase was detected in the SW480 cells. The expressions of TFAM protein were obviously increased in the SW480, HT-29, and HCTll6 cells as compared with the normal colon cell strain （P value was 0.000 2, 0.003 8, and 0.001 6, respectively）, among which the most notable increase was detected in the SW480 cells. After up-regulating TFAM by plasmid transfection, the proliferation of the pcDNA3.1-TFAM-SW480 cell was increased significantly as compared with the pcDNA3.1-SW480 cell at 96 h and 120 h after transfection by the MTT test （P〈0.000 1）. The proliferation of the pcDNA3.1-TFAM-SW480 cell was increased significantly as compared with the pcDNA3.1-SW480 cell at 48 h after transfection by the BrdU test （P〈0.001 0）. Conclusion TFAM expression is high in colon cancer. Up-regulated TFAM could promote the proliferation of colon cancer cells.
出处
《中国普外基础与临床杂志》
CAS
2015年第5期576-580,共5页
Chinese Journal of Bases and Clinics In General Surgery
关键词
线粒体转录因子A
结肠癌
增殖调控
Mitochondrial transcription factor A
Colon cancer
Proliferation regulation
