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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 investigate the effects of trichlorethylene (TCE) on the expression of three cytochrome P450 enzyme genes (CYP1A1, CYP2E1, CYP3A4) in human lymphocytoma cell line (MCL 5) and investigate the relationship between dose response and time response relationship. Methods Cells were treated with 0, 5, 10, 15 and 20 mmol / L LTCE for 12, 24, 48 and 72h respectively using conventional cell culture methods. CDNA was synthesized using a kit of purified RNA and cDNA, then three CYP450 genes were expressed by reverse transcription polymerase chain reaction (RT PCR) and β Actin was used as an internal control to analyze the intensity of gene expression at different doses and times. The results showed that the expression of CYP1A1 was up-regulated in LTMCs treated with LTCE for 48 hours, and the up-regulation tendency was enhanced with the prolongation of treatment time. The expressions of CYP2E1 and CYP3A4 Not affected by the length of TCE processing. The expression of three kinds of CYP450 enzyme genes was influenced by the dose of TCE, 3 showed a tendency of up-regulation with the increase of 0 5 mmol L, 10, 15, 20 mmol L dose. Conclusion TCE has obvious induction effect on CYP2E1, CYP1A1 and CYP3A4 genes in CYP450 enzyme system. The results of these genes induced may lead to the increase of corresponding enzyme activity, at the same time strengthen the metabolism of TCE, Metabolites increase.