CYP1A2基因在三氯乙烯毒性中的作用研究  

Role of CYP1A2 gene in trichlorethylene toxicity

作　　者：徐新云[1] 毛侃琅[1,2] 彭朝琼[1] 吴德生[1] 周丽[1] 秦逍云[1,2] 谭琴[1,2] 

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,深圳518055 [2]深圳大学生命科学学院,深圳518060

出　　处：《职业卫生与应急救援》2013年第3期115-118,共4页Occupational Health and Emergency Rescue

基　　金：深圳市科技计划重点项目(编号:201101015)

摘　　要：[目的]应用分子克隆技术,建立CYP1A2基因沉默细胞株和CYP1A2基因高表达细胞株,研究CYP1A2基因对三氯乙烯毒性的影响。[方法]将三氯乙烯分别染毒L02肝细胞、CYP1A2基因沉默细胞和CYP1A2基因高表达细胞,染毒剂量分别为0.25 mmol/L、0.50 mmol/L、1.00 mmol/L、2.00 mmol/L和4.00 mmol/L,以DMSO作为对照溶剂,染毒12 h,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)表达水平。[结果]三氯乙烯染毒CYP1A2基因沉默细胞后,Bcl-2的mRNA表达水平高于对照组(P<0.01),三氯乙烯部分剂量组凋亡基因Caspase-3、Caspase-8、Caspase-9和癌基因c-fos、c-myc表达水平低于对照组(P<0.05或P<0.01)。三氯乙烯染毒CYP1A2基因高表达细胞后,Bcl-2的mRNA表达水平相比对照组下降(P<0.01),Caspase-3、Caspase-8和Caspase-9 mRNA表达水平相比对照组升高(P<0.01);CYP1A2高表达细胞中癌基因c-fos、c-myc、k-ras表达水平高于对照组(P<0.01),P53表达水平低于对照组(P<0.01)。[结论]在体内代谢与CYP1A2存在密切关系,沉默CYP1A2基因可以减少三氯乙烯在肝细胞内代谢活化,导致三氯乙烯对凋亡基因和癌基因表达水平下降;三氯乙烯在CYP1A2基因高表达细胞中,表现出对凋亡基因和癌基因的促进表达作用。[Objective]To study the role of CYPIA2 gene in trichlorethylene （TCE） toxicity by molecular cloning technology. [ Methods ] CYPIA 2-silenced hepatocytes and hepatocytes with CYPIA2 gene over expression have been previously constructed by our laboratory. The normal hepatocytes （L02 cells）, CYPIA 2-silenced hepatocytes and hepatocytes with CYPIA2 gene over expression were treated with TCE for 12 h at doses of 0.25, 0.50, 1.00, 2.00 and 4.00 mmol/L,then the expression of apoptosis genes （Bcl-2,Caspase-3,Caspase-8,Caspase-9） and oncogenes （c-fos,c-myc,k-ras^P53）was observed. [Results]After TCE treatment,Bcl-2 expression levels in CYPIA2-silenced hepatocytes increased when compared with that of control （P 〈 0.01 ）, while the mRNA expression levels of apoptosis genes （ Caspase-3, Caspase-8, Caspase-9） and oncogenes （c-fos, c-myc ） decreased in CYPIA2-silenced hepatocytes compared with that in control （P 〈 0.05 or P 〈 0.01）. Additionally,Bcl-2 levels were significantly lower in CYPIA 2-overexpressed hepatocytes than that of control（P 〈 0.01 ）, the expression levels of apoptosis genes （ Caspase-3 , Caspase-8 , Caspase-9 ）and oneogenes （ c-fos ,c-myc ,k-ras ）increased significantly in CY PIA 2-overexpressed hepatocytes when compared with the control,but P53 levels were significantly lower than that of the control （P 〈 0.01）. [ Conclusion ] These findings indicated that CYPIA2 plays an important role in TCE metabolism in vivo. CYPIA 2 gene silencing could reduce TCE activation in hepatocytes,subsequently lead to decrease of apoptosis genes and oncogenes expression treatment. However, CYPIA2 gene over expression seems to promote apoptosis genes and oncogenes expression after TCE treatment.

关 键 词：三氯乙烯 肝细胞 CYP1A2 CYP1A2基因沉默 CYP1A2基因高表达 凋亡基因 癌基因 

分 类 号：R551[医药卫生—血液循环系统疾病] Q786[医药卫生—内科学]