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目的 在体外人神经细胞瘤细胞株 (SK N AS)中 ,以二恶英 (TCDD)、三甲基胆蒽 (3 MC)作为诱导化合物 ,探讨多环芳烃受体 (AHR、ARNT)基因与细胞色素P45 0 1A1/1B1(CYP1A1、CYP1B1)基因表达的调控 ,并研究其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 ,用二甲基亚砜 (DMSO)、5 0、5 0 0、10 0 0nmol/LTCDD和 0 1、1 0、10 0μmol/L 3 MC处理细胞 12h、2 4h、48h、72h ,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反应(RT PCR)表达AHR、ARNT和CYP1A1、CYP1B1基因 ,以 β actin作为内对照 ,分析不同处理剂量、时间时基因表达的强度。结果 4种基因在SK N AS中都有基本的表达 ,TCDD在 5 0 0nmol/L以上 ,染毒处理 48h、72h ,对AHR、ARNT及CYP1A1基因表达都有上调作用 ;5 0nmol/L组在染毒处理 72h后对AHR基因表达 ,5 0 0和 10 0 0nmol/L组分别在染毒处理 72h和 48h时对CYP1B1基因表达也有上调作用 ;3 MC 1 0、10 0 μmol/L在染毒 72h后对AHR、ARNT、CYP1A1基因表达有上调作用 ,其中 10 0 μmol/L组在染毒 48h后就有上调作用 ,但对CYP1B1基因表达上调仅仅在 10 0 μmol/L染毒 48h组。结论 本研究结果提示 ,TCDD、3 MC两种多环芳烃类化合物在SK N AS中 ,在一
OBJECTIVE: To investigate the expression of polycyclic aromatic hydrocarbon receptor (AHR, ARNT) genes in vitro and in vitro using dioxygenin (TCDD) and trimethycholanthrene (3 MC) as inducing compounds Cytochrome P45 0 1A1 / 1B1 (CYP1A1, CYP1B1) gene expression regulation, and study the relationship between dose response and time response. Methods The cells were treated with DMSO (DMSO), 5 0, 500, 10 000 nmol / LTCDD and 0 1, 10, 10 μmol / L 3 MC for 12 h, 24 h, 48h and 72h, cDNA was synthesized by using kit of purified RNA and cDNA synthesis. AHR, ARNT, CYP1A1 and CYP1B1 were expressed by reverse transcriptase polymerase chain reaction (RT PCR) and β actin was used as internal control. The intensity of gene expression at time. Results The four kinds of genes showed basic expression in SK N AS. TCDD was above 500 nmol / L, and 48 h and 72 h after exposure. The expressions of AHR, ARNT and CYP1A1 were all up-regulated. The expression of AHR gene was up-regulated after 72h exposure and the expression of CYP1B1 gene was also up-regulated at 50 and 100 nmol / L groups at 72h and 48h, respectively. After 72h, the expression of AHR, ARNT and CYP1A1 genes were up-regulated. The up-regulation of CYP1B1 gene expression was observed at 48 h after exposure to 10 μmol / L, but only up to 48 h after exposure to 100 μmol / L CYP1B1. Conclusion The results of this study suggest that TCDD, 3 MC two polycyclic aromatic hydrocarbons in the SK N AS, in a