用cDNA-AFLP技术筛选新生牛软骨无血管区的高表达基因

Screening High Expressed Genes in Avascular Region of Cow Cartilage by cDNA-AFLP Analysis

软骨一直被认为是内源性血管生成抑制物的重要来源.为更好理解软骨血管生成抑制的遗传学基础,对软骨中与血管生成抑制有关的基因进行了初步的探索.本研究应用cDNA-AFLP技术对新生牛关节骨骺软骨无血管区和有血管区的差异表达基因进行分析,筛选无血管区特异表达或高表达的基因,共得到43个无血管区高表达的片段.通过对片段进行亚克隆、测序及同源性分析,结果发现,16个片段与已知基因同源,25个片段只在牛的基因组数据库中找到同源序列,另有2个片段未找到同源序列.在同源基因中,肌钙蛋白Ⅰ亚型其血管生成抑制作用已有较详细报道;S100A7与血管生成和肿瘤发生的关系也已有阐述.另外14个同源基因按生物过程分析,发现参与了细胞周期调节、信号传递、细胞间相互作用及新陈代谢等多个生物过程.研究结果提示,牛软骨组织无血管区的无血管状态可能由多基因介导,其中一些可能是已知或未知的新基因.

Abstract The mammalian cartilage has been known as an important resource of angiogenic inhibitors for a long time. So, pilot studies were conducted for better understanding the genetic basis of angiogenic inhibition transcript derived fragments （TDFs） were identified as high expressed ones at the avascular region of cartilage by 90 primer combinations. Subsequent to subcloning, sequencing and homologously analyzing, it was found that, among them, 16 sequences were homologous with the known genes, and 25 sequences were found homology in cow genome database but no genes or ESTs, and the other 2 sequences had no significant hits to sequences currently in public databases. The homologous gene of troponin I had been reported to have the function of anti-angiogenesis, and SI00A7 had been found to be relative to cancer formaton. The other homologous genes were found to involve in transcription, cell cycle, signal transduction mechanism, cellular transport and metabolism etc. The finding suggested that the angiogenic inhibition of cartilage be controlled by multiple-genes, and many of which might belong to novel genes.