南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (11): 1548-1551.

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基因芯片筛选门静脉高压症脾亢脾和正常脾巨噬细胞中差异表达基因

闫峰； 李威； 陈君填； 曾永明； 郭毓文； 章斐然； 李宗芳；   

  1. 厦门大学附属中山医院博士后工作站； 汕头大学医学院附属第一医院普通外科； 西安交通大学第二医院普通外科 福建厦门361004； 广东汕头515041； 陕西西安710004；
  • 出版日期:2006-11-20 发布日期:2006-11-20
  • 基金资助:
    国家自然科学基金(30170909)~~

cDNA microarray-based screening of differentially expressed genes in macrophages in the spleen of patients with portal hypertension and hypersplenism

YAN Feng1,LI Wei2,CHEN Jun-tian2,ZENG Yong-ming2,GUO Yu-wen2,ZHANG Fei-ran2,LI Zong-fang3 1Postdoctoral Station,Zhongshan Hospital Affiliated to Shantou University,Shantou 361004,China;2Department of General Surgery,First Affiliated Hospital,Medical College of Shantou University,Shantou 515041,China;3Department of General Surgery,Second Hospital of Xi’an Jiaotong University,Xi’an 710004,China   

  1. 厦门大学附属中山医院博士后工作站； 汕头大学医学院附属第一医院普通外科； 西安交通大学第二医院普通外科 福建厦门361004； 广东汕头515041； 陕西西安710004；
  • Online:2006-11-20 Published:2006-11-20

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摘要： 目的运用基因芯片技术检测门静脉高压症脾亢脾和正常脾巨噬细胞中基因表达的差异,探讨其在门静脉高压症脾亢发生中的意义。方法提取门静脉高压症脾亢脾巨噬细胞和正常脾巨噬细胞的总RNA,分别用标有荧光素的dCTP反转录制备cDNA探针,将探针与含有14112点cDNA的Biostar-H140scDNA表达谱芯片杂交后扫描荧光强度。上述方法重复3次,从而筛选出恒定的差异表达基因。结果3张芯片分别检测到896、1330和898个差异表达基因,恒定的差异表达基因共有121个,占总基因数的0.86%。其中表达上调的已知基因有21个,表达下调的已知基因有73个。差异表达基因涉及离子通道和运输蛋白、细胞周期蛋白类、细胞骨架和运动、细胞受体、细胞信号和传递蛋白、代谢、免疫相关等多个方面。这些基因可能与门静脉高压症脾亢的发病机制有关。结论采用基因芯片技术筛选门静脉高压症脾亢脾和正常脾巨噬细胞中差异表达的基因,可能为其病因和发病机制的研究提供新思路。 

Abstract: Objective To identify the differentially expressed genes associated with hypersplenism in patients with portal hypertension.Methods The total RNA were extracted from the macrophages isolated from normal spleen and the spleen of patients with portal hypertension and reversely transcribed to cDNA with the incorporation of fluorescent(cy3 and cy5)-labeled dCTP to prepare the hybridization probes.After hybridization of Biostar-H140s chip containing 14 112 spots of cDNAs with the prepared probes,the gene chip was scanned for fluorescence intensity to screen the differently expressed genes.Three gene chips were used for hybridization and only the genes with differential expression in all the three chips were considered to associate with hypersplenism in patients with portal hypertension.Results Totaling 896,1330 and 898 genes were identified to be differentially expressed by the three chips,respectively,and 121 genes(0.86%) showed differential expression in all the three chips,including 21 up-regulated known genes and 73 down-regulated known genes.The differently expressed genes were functionally related with ion channels and transport proteins,cyclins,cytoskeleton,cell receptors,cell signal transduction,metabolism,immunity,and so forth.These genes might be involved in hypersplenism in the condition of portal hypertension.Conclusion cDNA microarray-based screening of differentially expressed genes in the macrophages in the spleen may provide new insights into the pathogenesis of hypersplenism in patients with portal hypertension. 

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