【摘要】 目的探讨二甲肼对大肠上皮细胞的作用和基因表达谱改变影响。方法以前期建立的永生化正常大肠上皮细胞为模型,采用软琼脂克隆形成实验探讨二甲肼的诱导转化作用,并用基因芯片技术揭示转化过程中的基因谱变化,实时荧光定量PCR和Western blot技术进行验证。结果经二甲肼作用后大肠上皮细胞在软琼脂中出现细胞克隆,与正常对照细胞比较,转化细胞有265个基因表达发生明显改变,包括148个上调基因和117个下调基因。分泌性白细胞蛋白酶抑制因子(secretory leukocyte protease inhibitor,SLPI)在转录和蛋白水平上均发生明显上调。结论二甲肼能诱导永生化正常大肠上皮细胞发生恶性转化,初步揭示了转化过程中基因表达谱的改变,为认识大肠癌发生的分子机制提供了有益线索。更多还原

【Abstract】 Objective To investigate the alteration of gene profile in transformation of colon epithelial cells by 1,2-dimethylhydrazine(DMH).Methods A conditionally immortalized normal human colorectal epithelial cell line was established and employed as a target for transformation by DMH.The alterations of gene expression profile was investigated by Genechip analysis and verified by quantitative real-time PCR and Western blotting.Results Cell colonies were shown in soft agar after DMH treatment.Compared to the normal cells,265 genes in the transformed cells significantly changed,including 148 up-regulated and 117 down-regulated.The mRNA and protein levels of secretory leukocyte protease inhibitor(SLPI) were all increased significantly.Conclusion This study demonstrates the alterations of gene expression profile in the transformation of colon epithelial cells by DMH,which may provide information for better understanding the molecular mechanism of colorectal carcinogenesis.更多还原