三氯乙烯对CYP2E1基因高表达细胞和正常肝细胞的毒性作用  

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

作　　者：徐新云[1] 毛侃琅[1] 袁建辉[1] 吴德生[1] 黄海燕[1] 秦逍云[1] 谭琴[1] 

机构地区：[1]深圳市疾病预防控制中心卫生毒理学重点实验室,深圳市现代毒理学重点实验室,518055

出　　处：《中华劳动卫生职业病杂志》2014年第10期723-727,共5页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：深圳市科技计划重点项目（201101015）

摘　　要：目的 应用分子克隆技术建立CYP2E1基因高表达细胞株,观察三氯乙烯（TCE）对CYP2E1基因高表达细胞和正常肝细胞的毒性,探讨CYP2E1基因在TCE毒性中的作用.方法 用不同剂量三氯乙烯（0、0.25、0.5、1.0、2.0和4.0 mmol/L）对CYP2E1高表达细胞和正常人肝细胞（L02细胞）进行染毒12h,观察凋亡基因（Bcl-2、caspase-3、caspase-8、caspase-9）和癌基因（c-fos、c-myc、k-ras、p53）表达变化.结果 TCE染毒引起L02肝细胞和CYP2E1高表达细胞Bcl-2表达水平下降,0.25～2.0 mmol/L TCE剂量时L02肝细胞Bcl-2表达下降50％～60％,但相同剂量组的CYP2E1高表达细胞Bcl-2表达水平明显高于正常肝细胞20％～50％; 0.5～4.0 mmol/L TCE剂量时CYP2E1高表达细胞中凋亡基因caspase-3、caspase-8、caspase-9表达水平比正常肝细胞高30％～600％,差异有统计学意义（P＜0.05或P＜0.01）.TCE染毒后对L02肝细胞和CYP2E1高表达细胞的癌基因表达水平有明显差异,在0.5 ～4.0 mmol/L TCE染毒时,CYP2E1高表达细胞的c-fos、k-ras和c-myc基因表达水平明显高于L02肝细胞,差异有统计学意义（P＜0.05或P＜0.01）,升高幅度达25％～120％; CYP2E1高表达细胞的p53表达水平比正常肝细胞表达下降10％～50％,差异有统计学意义（P＜0.05或P＜0.01）.结论 TCE对正常肝细胞和CYP2E1高表达细胞凋亡基因和癌基因表达存在明显差异,提示CYP2E1是TCE在体内代谢的重要因素,与TCE毒性存在一定关系.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 PC R.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.

关 键 词：三氯乙烯 肝细胞 细胞色素P-450 基因表达 

分 类 号：R114[医药卫生—卫生毒理学]