利用表达谱基因芯片筛选溃疡性结肠炎差异表达基因

目的:利用人类全基因组表达谱芯片技术,分析溃疡性结肠炎患者和健康者基因表达谱差异,筛选出溃疡性结肠炎相关基因。方法:采用Trizol法提取8例溃疡性结肠炎患者和8例健康对照者结肠粘膜组织总RNA并纯化,逆转录合成c DNA,利用荧光染料Cy3标记aa UTP,转录合成标记的c RNA,并与Agilent人类全基因组表达谱芯片杂交,扫描荧光信号图像,对芯片原始数据进行归一化处理,利用倍数差异和t检验计算筛选出相关差异表达基因,采用DAVID在线分析系统进行基因的功能注释和关联分析,明确差异基因的生物学功能,并对部分差异表达基因进行实时荧光定量PCR验证。结果:筛查出溃疡性结肠炎结肠粘膜组织差异表达基因4132个,其中上调基因2004个,下调基因2128个。选取6条差异表达基因进行PCR验证,结果有3条基因表达上调,3条基因表达下调,表达趋势与芯片结果一致。结论:溃疡性结肠炎患者与健康对照者基因表达存在明显差异,分析这些差异表达基因有助于我们探索溃疡性结肠炎的发病机制,为疾病的治疗提供理论依据。

Identification of Differential Expression Genes Related to Ulcerative Colitis by Gene Expression Profiles Chip

Objective:To detect and compare the gene expression profile between ulcerative colitis patients and healthy control using cDNA microarray method and screen differential expression genes related with UC.Methods:Total RNA was extracted and purified from 8 UC patients and 8 controls. The first and second cDNA strains were generated and transcribed into cRNA, fluorescence dye Cy3 was used to label the cRNA. The labeled cRNA was used to hybrid with the Agilent expression profile chip, the fluorescence signal was scanned by the computer and the original data was normalized. The differential expression genes was screened by t-test and identified the biological function of genes by the DAVID online analysis system. Parts of these genes were analyzed by real-time PCR.Results:4132 genes were differentially expressed in gene profile of UC, 2004 were up-regulated and 2128 down-regulated. The expression trend of six genes for further verification was consistent with the array(3 up-regulated and 3 down-regulated).Conclusion:Distinctly different expression profiles exist in UC patients and healthy control, analyzing these genes may help clarify the pathogenesis of UC and provide the theoretical basis for the treatment of diseases.