用cDNA微阵列芯片检测苯中毒细胞色素P450基因表达差异

Analysis on differential expression profile of cytochrome P450 genes in benzene poisoning cells using cDNA microarray

目的利用cDNA芯片技术检测苯中毒细胞色素P 450的基因表达谱差异。方法选取7名诊断为不同程度接苯中毒的工人(疑似1例、轻度2例、中度2例和重度2例),并设年龄、工龄匹配的健康者做对照,分离外周血白细胞,用T rizo l试剂盒抽提总RNA,纯化、逆转录生成cDNA产物,分别用荧光染料Cy3和Cy5标记后与含32条细胞色素P 450基因的微阵列芯片进行杂交。结果共有6条细胞色素P 450基因呈差异性表达,包括CYP 4F 3、CYP 1A 1、CYP 27A 1、CYP 1B 1、CYP 2B 6、CYP 51,其中CYP 4F 3基因在所有苯中毒工人中均表达上调。结论苯中毒工人细胞色素P 450基因的表达异常可能在苯诱导的血液毒性中起重要作用。

Objective To detect differential expression of cytochrome P450 genes in patients diagnosed with benzene poisoning of different degrees by cDNA microarray. Methods Peripheral white blood cells were isolated from seven patients diagnosed with benzene poisoning of different degrees and seven age-and sexmatched normal subjects were selcted. Total RNA was extracted and purified, followed by reverse transcription to cDNAs with concomitant incorporation of fluorescent dCTP （Cy3 or CyS）. The cDNAs were used as probes in microarray. Fluorescent signals were scanned to detect cytochrome P450 genes differentially expressed in patients and normal subjects. Results Among 32 cytochrome P450 genes, six genes （CYP4F3, CYP1A1, CYP27A1, CYP1B1, CYP2B6, CYP51） were found to be differentially expressed between the cells of benzene-poisoned patients and normal controls cells, CYP4F3 gene was up-regulated among all patients. Conclusion Differentially expressed cytochrome P450 genes in benzene poisoning patients possibly play important role in benzene-induced hematotoxicity, cDNA microarray technique is useful to indicate expression mode of benzene poisoning.