摘要
目的 探讨三氯乙烯 (Trichloroethylene ,TCE)对人体淋巴细胞瘤细胞株 (MCL 5 )中 3种细胞色素P4 5 0酶基因 (CYP1A1、CYP2E1、CYP3A4 )表达的影响 ,并研究剂量反应关系和时间反应关系。方法 用常规的细胞培养方法 ,0 5、1 0、1 5、2 0mmol LTCE处理细胞 12、2 4、4 8、72h。利用提纯RNA和合成cDNA的药盒 ,合成cDNA ,然后逆转录聚合酶反应 (RT PCR)表达 3种CYP4 5 0基因 ,以β Actin作为内对照 ,分析不同处理剂量和时间时基因表达的强度。结果 在MCL 5细胞株中都有基本的表达 ,CYP1A1表达在用 1 0、1 5、2 0mmol LTCE处理 4 8h后有被上调的趋势 ,而且上调趋势随处理时间延长而加强 ;CYP2E1、CYP3A4表达不受TCE处理时间长短的影响。 3种CYP4 5 0酶基因的表达受TCE剂量的影响 ,3随 0 5mmol L ,1 0、1 5、2 0mmol L剂量的增加有上调的趋势。结论 TCE对CYP4 5 0酶系统中的CYP2E1、CYP1A1、CYP3A4基因有明显的诱导作用 ,这些基因被诱导后的结果 ,可能会导致相对应酶活性的增加 ,同时加强对TCE的代谢 ,使TCE的代谢产物增加。
Objective To study the regulation of CYP1A1,CYP2E1,CYP3A4 genes expression in the MCL-5 cell line after treatment with trichloroethylene (TCE) and its relationships of dose-response and time-response.Methods The human cell line MCL-5 was treated with DMSO,0.5、1.0、1.5、2.0 mmol/L TCE for 12、24、48、72 h respectively.RNA of cell line was purified by RNA kit.The cDNA synthesis kit was used to synthesize cDNA.Then RT-PCR was conducted to amplify the fragment of CYP1A1,CYP2E1,CYP3A4 DNA.Results CYP1A1,CYP2E1,CYP3A4 genes were constitutively expressed.CYP1A1 was induced after 48 h treated with 1.0、1.5、2.0 mmol/LTCE,respectively,and this induction at three doses was time-dependent.The induction of CYP2E1,CYP3A4 was not time-response.Otherwise,the induction of CYP1A1,CYP2E1,CYP3A4 genes was concentration-dependent on TCE.The up-regulation of CYP1A1,CYP2E1,CYP3A4 was positively correlated with TCE doses.Conclusion The results showed that the expression of CYP1A1,CYP2E1,CYP3A4 could be significantly induced by TCE.The MCL-5 cell line might be a good model for further study on CYP450 expression.
出处
《卫生毒理学杂志》
CSCD
北大核心
2001年第3期140-143,共4页
Journal of Health Toxicology
