摘要
目的 :研究人类退变椎间盘的基因表达谱 ,分析人退变椎间盘基因表达水平的变化。方法 :按条件优化的一步法抽提人退变及正常椎间盘组织的总RNA各 3例 ,分别用cy3 ,cy5荧光标记 ,获得 2组椎间盘cDNA的探针 ,与含有 40 96条人类全长基因的cDNA表达谱芯片杂交 ,扫描芯片荧光信号图像 ,对所获得的基因进行生物信息学分析。结果 :在 40 96条基因中 ,有差异表达的基因 70 6条 ,3 5 8条基因表达量明显下降 ,2 98条基因表达量明显上升。在有差异表达的基因中 ,细胞凋亡相关类蛋白 8条 ,其中上皮细胞膜蛋白 (EMP 1 )基因的表达上调明显。结论 :退变椎间盘的基因表达发生变化 ,细胞凋亡增加可能是椎间盘退变发生和发展的因素之一。
Objective:To investigate the gene expression changes in normal and degenerated lumbar intervertebral disc in humans,providing information for clinical.Methods: The PCR products of 4 096 human genes were spotted onto a chemical-material-coated-glass slides. The total RNAs were isolated from the tissues. Both the mRNAs from the degenerated and normal lumbar intervertebral disc in humans were reversely transcribed to the cDNAs. The cDNAs with the incorporations of fluorescent dUTP was used for preparing the hybridization probes. The mixed probes were then hybridized to the cDNA microarray. After high-stringent washing,the cDNA microarray was scanned for the fluorescent signals and analyzed by computer image analysis. Results:Among the 4 096 targets,there were 706 genes whose expression levels differed between the degenerated and normal lumbar intervertebral disc in all 6 cases,including 298 up-regulated and 358 down-regulated ones. Conclusion:DNA microarray technology is an effective technique in screening for differentially expressed genes between the degenerated and normal lumbar intervertebral disc. Cell apoptosis may play a role in the process of lumbar intervertebral disc degeneration.
出处
《解剖学杂志》
CAS
CSCD
北大核心
2004年第4期348-351,F002,共5页
Chinese Journal of Anatomy
