论文部分内容阅读
目的 探讨四氯化碳 (carbontetrachloride ,CCl4)和乙醇 (ethanol ,EtOH)对人体淋巴细胞瘤细胞株 (MCL 5 )中 3种细胞色素P45 0酶基因 (CYP1A1、CYP2E1、CYP3A4)表达的影响 ,并研究剂量与效应的关系。方法 用常规的细胞培养方法 ,用DMSO ,0 0 5、0 10、0 15、0 2 0mmol/LCCl4和 5 0、10 0、15 0、2 0 0mmol/LEtOH处理细胞 48h ,利用提纯RNA和合成cDNA的药盒 ,合成cDNA ,然后通过逆转录聚合酶链反应 (RT PCR)表达 3种CYP45 0酶基因 ,以 β Actin作为内对照 ,分析不同处理剂量时基因表达强度。结果 3种CYP45 0酶基因在MCL 5细胞株中都有基本表达 ,CYP2E1和CYP3A4表达在用 0 0 5、0 10、0 15、0 2 0mmol/LCCl4和 5 0、10 0、15 0、2 0 0mmol/LEtOH处理细胞 48h后有被上调的趋势。结论 本研究结果提示 ,CCl4和EtOH对CYP45 0酶系统中的CYP2E1、CYP3A4基因有明显的诱导作用 ;这些基因被诱导后 ,可能会导致相对应酶活性的增加 ,同时加强对CCl4、EtOH的代谢 ,使他们的毒性代谢产物增加。
Objective To investigate the effects of carbon tetrachloride (CCl4) and ethanol (EtOH) on the expression of three cytochrome P45 0 genes (CYP1A1, CYP2E1, CYP3A4) in human lymphoma cell line MCL 5 Study the relationship between dose and effect. Methods The cells were treated with DMSO, 0 05,0 10,0 15,0 2 0 mmol / L HCl and 50, 100, 0 0, 0 0 0 0 mmol / LEtOH for 48h. cDNA kit, cDNA was synthesized, and then three kinds of CYP45 0 enzyme genes were expressed by reverse transcription polymerase chain reaction (RT PCR). β Actin was used as an internal control to analyze the gene expression intensity at different treatment doses. Results The three CYP45 0 genes were expressed basically in MCL 5 cell lines. The expressions of CYP2E1 and CYP3A4 were significantly different between the cells treated with 0 0 5,0 10,0 15,0 2 0 mmol / L HCl and 50 0,10 0,15 0,2 0 0mmol / LEtOH treated cells 48h after the trend was up. Conclusions The results of this study suggest that CCl4 and EtOH may induce the CYP2E1 and CYP3A4 genes in the CYP45 0 enzyme system. The induced induction of these genes may result in the increase of the corresponding enzyme activity and the enhancement of the metabolism of CCl 4 and EtOH , To increase their toxic metabolites.