[三氯乙烯毒性对正常人类肝细胞及CYP2E1基因过表达肝细胞的影响]

[Effects of trichloroethylene toxicity on normal human liver cells and hepatocytes with CYP2E1 gene overexpression].

作者信息

Xu Xinyun, Mao Kanlang, Yuan Jianhui, Wu Desheng, Huang Haiyan, Qin Xiaoyun, Tan Qin

机构信息

Shenzhen Center for Disease Control and Prevention, Shenzhen key laboratory of health toxicology, Shenzhen key laboratory of modern toxicology, Shenzhen, Guangdong 518055, China.

出版信息

Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2014 Oct;32(10):723-7.

PMID:25533359

Abstract

OBJECTIVE

To investigate the effects of trichloroethylene (TCE) toxicity on the normal human liver cells (L02 cells) and hepatocytes with CYP2E1 gene overexpression which was constructed through molecular cloning technology in our laboratory, then to explore the roles of CYP2E1 gene in TCE toxicity.

METHODS

L02 cells and hepatocytes with CYP2E1 overexpression were treated with various doses of TCE (0,0.25, 0.5, 1.0, 2.0, 4.0 mmol/L) for 12h, the expression of apoptosis genes (Bcl-2, Caspase-3, Caspase-8, Caspase-9) and oncogenes (c-fos, c-myc, k-ras, p53) were determined by real-time fluorescent PCR.

RESULTS

Bcl-2 mRNA expression levels increased significantly in normal liver cells and CYP2E1-overexpressing cells after TCE treatment, Bcl-2 levels were 20%∼50%higher in CYP2E1-overexpressing cells than in L02 liver cells at doses of 0.25∼2.0 mmol/L TCE. Caspase-3, Caspase-8 and caspase-9 mRNA expression increased by 30%∼600% in CYP2E1-overexpressing cells at doses of 0.5∼4.0 mmol/L TCE when compared with L02 cells (P < 0.01). Additionally, c-fos, k-ras and c-myc mRNA expression levels were 25%∼120% higher in CYP2E1-overexpressing cells than in L02 cells (P < 0.01), p53 mRNA expression levels were lower 10%∼50% in CYP2E1-overexpressing cells than in L02 cells (P < 0.05 or P < 0.01).

CONCLUSIONS

There were significant differences for apoptosis gene and oncogene expression levels between normal liver cells and CYP2E1-overexpressing cells after they were treated with TCE, these findings indicated that CYP2E1 might play an important role in TCE metabolism in vivo.

摘要

目的

研究三氯乙烯（TCE）毒性对正常人类肝细胞（L02细胞）以及本实验室通过分子克隆技术构建的CYP2E1基因过表达的肝细胞的影响，进而探讨CYP2E1基因在TCE毒性中的作用。

方法

用不同剂量的TCE（0、0.25、0.5、1.0、2.0、4.0 mmol/L）处理L02细胞和CYP2E1过表达的肝细胞12小时，通过实时荧光定量PCR检测凋亡基因（Bcl-2、Caspase-3、Caspase-8、Caspase-9）和癌基因（c-fos、c-myc、k-ras、p53）的表达。

结果

TCE处理后，正常肝细胞和CYP2E1过表达细胞中Bcl-2 mRNA表达水平显著升高，在0.25至2.0 mmol/L TCE剂量下，CYP2E1过表达细胞中的Bcl-2水平比L02肝细胞高20%至50%。与L02细胞相比，在0.5至4.0 mmol/L TCE剂量下，CYP2E1过表达细胞中Caspase-3、Caspase-8和Caspase-9 mRNA表达增加了30%至600%（P < 0.01）。此外，CYP2E1过表达细胞中c-fos、k-ras和c-myc mRNA表达水平比L02细胞高25%至120%（P < 0.01），CYP2E1过表达细胞中p53 mRNA表达水平比L02细胞低10%至50%（P < 0.05或P < 0.01）。