用基因芯片体外研究苯暴露大鼠骨髓细胞基因的表达

A Study of Gene Expression Patterns in Rat's Bone Marrow Cells Exposed to Benzene with Genechip in Vitro

目的 研究正常和苯处理后的大鼠骨髓细胞的基因表达谱 ,大规模分析苯作用后骨髓细胞基因表达水平的变化。方法 1实验大鼠骨髓细胞分为两组 ,一组为正常对照组 ,一组为染苯组 ;2从两组骨髓细胞中抽提总 RNA ,经反转录分别用 Cy3、Cy5荧光标记 ,获得 c DNA探针 ;3c DNA探针与基因谱芯片杂交 ,结果由扫描仪扫描并用软件进行分析统计。结果 骨髓细胞暴露苯后 2 0 0条 (4 .88%)基因发生明显变化 ,其中 1 5 4条 (77%)基因已在 Gene Bank中登录。这 2 0 0条基因中 ,有 94条基因表达降低 (Cy5 / Cy3* <0 .5 ) ,1 0 6条基因表达升高 (Cy5 / Cy3* >2 .0 )。结论 利用基因芯片技术结合体外细胞培养能大规模地研究苯毒性作用的基因表达谱 ,初步筛选出苯毒性作用相关基因。对进一步阐明苯毒性在基因水平的发病机理有十分重要的作用。

Objective To study the gene expression patterns of rat's bone marrow cells(BMC) after benzene exposure.Methods ① The BMC was divided into 2 groups: control group and benzene exposure group;② Total RNA from BMC was extracted, the probes were prepared by labeling normal and benzene exposed to total RNA with Cy3 dCTP and Cy5 dCTP separately through reverse transcription;③ The arrays were hybridized against the cDNA probe mixture and the fluorescent signals were scanned, the obtained data were analyzed with software.Results Significant changes were observed in 200 genes(4.88%),in which 154 genes(77%) have been registered in Genebank. The average ratio between Cy5 and Cy3 signals of 94 genes were lower than 0.5,while the average ratio between Cy5 and Cy3 signals were higher than 2.0.Conclusion Using gene microarray and cell cluture in vitro the gene expression patterns induced by benzene and the benzene toxic related genes can be studied.The method plays an important role in interpreting benzene toxic mechanism at gene levels.