魏妮 郭长存 沙素梅 徐斌 白槟 余鹏飞 吴开春.利用表达谱基因芯片筛选溃疡性结肠炎差异表达基因[J].,2015,15(1):53-57 |
利用表达谱基因芯片筛选溃疡性结肠炎差异表达基因 |
Identification of Differential Expression Genes Related to UlcerativeColitis by Gene Expression Profiles Chip |
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DOI: |
中文关键词: 溃疡性结肠炎 基因表达谱芯片 差异表达基因 |
英文关键词: Ulcerative colitis Gene expression profiles chip Differential expression genes |
基金项目:国家自然科学基金项目(43411C1452) |
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中文摘要: |
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cDNA,利用荧
光染料Cy3标记aaUTP,转录合成标记的cRNA,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。 |
英文摘要: |
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. |
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