叶绿酸抑制细胞恶性转化的基因表达分析

Study on gene expression in cells treated anti-transformationally with chlorophyllin

目的 采用cDNA微阵列技术探索人支气管上皮细胞经叶绿酸抗转化处理后基因的差异表达。方法 人支气管上皮细胞株 1 6HBE分别经二羟环氧苯并芘 (BPDE)单独处理及叶绿酸与BPDE联合处理后 ,提取 2种细胞的总RNA ,逆转录合成cDNA并以Cy3 -dCTP和Cy5 -dCTP荧光素分别标记制作探针。探针混合后与含 4 0 0 0个人类基因的芯片杂交。以ScanArray 40 0 0扫描仪扫描芯片 ,以GenPixPro 3 0软件分析荧光信号 ,然后对差异表达的基因进行生物学信息分析。结果 叶绿酸与BPDE联合处理的细胞样与BPDE单独处理的细胞样比较呈显差异表达的基因有 1 0 6个 ,其中 67个 (63 9% )下调 ,另 39个 (36 8% )上调。结论 叶绿酸的抗转化活性与应激反应基因 ,免疫相关基因 ,DNA合成和修复基因 。

Objective Using cDNA microarray to screen for the differentially expressed genes in human bronchial epithelial cell line 16HBE after anti-transforming treatment with chlorophyllin.Methods 16HBE cells were treated with dihydroxyepoxy benzp pyrene(BPDE)only and chlorophllin with BPDE jointly.The total RNAs were extracted from these cells.The cDNA probes were prepared by labeling with Cy3-dCTP and Cy5-dCTP respectively through reverse transcription.The mixed probes were then hybridized to the cDNA microarray chips containing 4000 human genes.The chips were scanned by ScanArray 4000 laser scanner.The acquired fluorescent signals were analyzed by GenPix Pro 3.0 software.Bioinformatical analysis of those differentially expressed genes was performed.Results 106 genes exhibited differential expression between cells treated with BPDE only and those with chlorophyllin and BPDE jointly.The expression of 67 genes(63.9%) was down-regulated and of 39 genes(36.8%) was up-regulated.Conclusion The regulation of genes including stress response genes,immune related genes,DNA synthesis and repairing genes,metabolic genes may be involved in antitransforming activity of Chlorophylin.