cDNA芯片法筛选压力负荷型心肌肥厚相关基因

Identification of genes related to mechanical load-induced cardiac hypertrophy by cDNA chip

应用cDNA芯片技术筛选腹主动脉绑扎后 4周大鼠的压力负荷型心肌肥厚相关基因。在反转录来自假手术及腹主动脉绑扎 (4周 )的大鼠心肌组织RNA的过程中 ,用α 3 3 P dATP进行探针的标记。然后将探针与微阵列尼龙膜杂交 ,高严谨度洗涤后分析杂交图谱。通过所获某种特定基因信号的相对强度值来比较其在对照及腹主动脉绑扎的大鼠心肌组织中表达水平的差异。结果显示有 2 35个基因片段在两组之间信号差别在 3倍以上 ,其中 5倍以上差异的基因有 5 5个 ,而在这 5 5个基因中有 2 6个基因功能未知。所得差异片段包括TGFβ受体III ,raf,ARF ,clk2等重要的信号传导分子与心肌肥厚的发生密切相关。

To isolate the gene fragments related to cardiac hypertrophy(4 weeks after arota constriction) by cDNA expression arrays analysis, the probe was synthesized by reverse transcribing RNA population using α- 33P-dATP.Each radioactively labeled probe was then hybridized to separate arrays. After a high-stringency wash, the hybridization pattern was analyzed. The relative expression levels of a given cDNA from two different RNA sources (one is from control group,a other one is from abdominal aorta-constriction group) was assessed by comparing the signal obtained. The results show that there were 235 fragments expressed differentially more than three times between normal and model group ,including 55 fragments expressed differentially more than five times,of which 26 were unknown genes. Some genes were closely correlated with the occurrence of cardiac hypertrophy including TGF-βIII receptor gene,raf oncogene, ARF, clk2,etc.