Dai Wentao, Chen Huajie, Yu Rian, He Lingfei, Chen Bing, Chen Xuemin
Department of Occupational and Environmental Health, Guangdong Key Lab of Molecular Epidemiology, School of Public Health, Guangdong Pharmaceutical University, Guangzhou 510310, China.
J Huazhong Univ Sci Technolog Med Sci. 2010 Dec;30(6):709-13. doi: 10.1007/s11596-010-0645-8. Epub 2010 Dec 22.
This study investigated the effect of cadmium on the telomerase activity, the expression of TERT, c-myc and p53 and the apoptosis of rat hepatocytes. The rats were administrated 5, 10 and 20 μmol/kg cadmium chloride intraperitoneally and sacrificed 48 h after the initial treatment. The telomerase activity of the rat hepatocytes was measured by the telomeric repeat amplification protocol (TRAP), and apoptosis was detected by flow cytometry. The mRNA expressions of TERT, c-myc and p53 were measured by reverse transcription-polymerase chain reaction (RT-PCR). C-myc and P53 proteins were determined by immunochemistry. The results showed that cadmium chloride increased the hepatocellular telomerase activity in a dose-dependant manner and induced the apoptosis of hepatocytes significantly. The value of relative coefficient between the telomerase activity and the apoptosis rate was 0.9398. RT-PCR revealed that specific bands corresponding to the TERT mRNA, c-myc mRNA, and p53 mRNA were displayed at 185, 342 and 538 bp respectively. Cadmium chloride could substantially increase the mRNA expressions of TERT, c-myc and p53 in rat hepatocytes, as compared with control. Moreover, cadmium chloride at the doses of 5, 10 and 20 μmol/kg could increase the content of P53 protein in rat hepatocytes obviously, but only that at the doses of 10 and 20 μmol/kg substantially promoted the c-myc protein level in rat hepatocytes. Our study herein suggested that cadmium may contribute to the carcinogenesis by activating telomerase, and overexpressing the mRNAs of TERT, c-myc and p53, and causing apoptosis of normal cells.
本研究调查了镉对大鼠肝细胞端粒酶活性、端粒酶逆转录酶(TERT)、c-myc和p53的表达以及细胞凋亡的影响。给大鼠腹腔注射5、10和20 μmol/kg的氯化镉,并在初次处理后48小时处死。通过端粒重复序列扩增法(TRAP)测定大鼠肝细胞的端粒酶活性,通过流式细胞术检测细胞凋亡。通过逆转录-聚合酶链反应(RT-PCR)测定TERT、c-myc和p53的mRNA表达。通过免疫化学法测定c-myc和P53蛋白。结果表明,氯化镉以剂量依赖的方式增加肝细胞端粒酶活性,并显著诱导肝细胞凋亡。端粒酶活性与凋亡率之间的相关系数值为0.9398。RT-PCR显示,对应于TERT mRNA、c-myc mRNA和p53 mRNA的特异性条带分别出现在185、342和538 bp处。与对照组相比,氯化镉可显著增加大鼠肝细胞中TERT、c-myc和p53的mRNA表达。此外,5、10和20 μmol/kg剂量的氯化镉可明显增加大鼠肝细胞中P53蛋白的含量,但只有10和20 μmol/kg剂量的氯化镉可显著提高大鼠肝细胞中c-myc蛋白水平。我们的研究表明,镉可能通过激活端粒酶、使TERT、c-myc和p53的mRNA过表达以及导致正常细胞凋亡而促进致癌作用。