用cDNA微阵列分析苯中毒肿瘤相关基因表达谱

Analysis on tumor related gene expression profiles in benzene poisoning using cDNA microarray

目的运用cDNA微阵列技术对苯中毒肿瘤相关基因表达谱进行分析研究。方法对7例接苯中毒工人及7例正常对照者全血中淋巴细胞分别进行RNA抽提,纯化后的mRNA配对进行逆转录制备杂交探针,利用7张含有2780个cDNA克隆的微列阵肿瘤相关基因芯片进行杂交,杂交信号用扫描仪进行扫描,然后将图像转化为基于荧光强度的数字信号,随后进行数据分析和处理。结果在7张肿瘤相关基因中共发现特异性差异表达基因44个,其中GRO1、TGFBR3、LYN等16个基因表达上调,FOSB、DJ1、MCT1等28个基因表达下调。结论苯暴露患者肿瘤相关基因的表达异常,提示其可能为致白血病的关键基因,在苯致白血病中起着重要的作用;采用基因芯片检测技术有利于全面揭示苯中毒肿瘤相关基因表达模式,快速高效地发现新的研究目标和基因治疗途径。

Objective To detect target genes for further study by way of analyzing the gene expression profiles of benzene poisoning by using cDNA microarray. Methods Peripheral mononuclear cells were isolated from seven patients with benzene poisoning of different degrees, and sevene age-and sex-matched normal subjects.Total RNA was extracted and purified, followed by revese transcription to cDNAs with concomitant incorporation of fluorescent dCTP（Cy3 or Cy5）.The cDNAs were used as probes in microarray of 2780 cloned eDNA. Fluorescent signals were scarmed to detect genes differentially expressed in patients and normal subjects. Results Among 7 pieces of eDNA microarray of 2780 tumour related genes, the expression of 16 genes, such as GRO1, TGFBR3, LYN ctc was upregulated, whereas the expression of 28 genes, such as FOSB, DJ- 1, MCT- 1 etc was down-regulated.Conclusion Abnormal expression of tumour related genes of patients exposed to benzene suggests that they may be the key genes, which play important role in benzene-lnduced leucocythemia, eDNA microarray technique is useful to indicate the expression mode of benzene poisoning tumour related genes, and to find rapidly and effectively new research object and the way of gene therapy.