目的应用表达谱芯片比较人强直性脊柱炎(ankylosing spondylitis,AS)组织与正常骶髂关节组织的基因表达差异,筛选AS疾病相关基因。方法选取本室采集的3例活动期AS标本为实验组,1例骨折标本为对照组。分别提取标本骶髂关节的滑膜组织与椎旁组织的RNA,制备探针后用于表达谱芯片杂交,筛选AS组织差异表达基因,并采用半定量RT-PCR法验证芯片结果。结果与正常组织样本相比,在AS组织中筛选获得差异表达基因237条,表达上调的已知基因91条,表达下调的已知基因78条;对其中20条基因进行半定量RT-PCR检测,结果与表达谱芯片结果一致。结论筛选出与AS相关的多个差异表达基因。
Objective To compare the gene expression difference between human ankylosing spondylitis(AS) and normal sacroiliac joint tissues,and to screen AS-associated genes via cDNA microarray.Methods Three AS samples in active phase were collected and used as a test group,and a fracture sample was used as a control group.Total RNA was isolated from synovial and paravertebral tissues of both AS and normal sacroiliac joints,and used to construct hybridization probes.The probes were hybridized to cDNA microarray GeneChip Human Genome U133 Plus 2.0 Array.The differentially expressed genes in AS tissues were identified.Finally,semi-quantitative RT-PCR was used to confirm the results of the cDNA microarray.Results Compared with the normal tissue sample,237 differentially expressed genes were detected in AS group,including 91 up-regulated and 78 down-regulated known genes.The semi-quantitative RT-PCR test revealed that the expression patterns of 20 candidate differentially expressed genes were consistent with the results of cDNA microarray.Conclusion A number of AS-associated differentially expressed genes are detected,and the present study will be helpful to unveil the molecular mechanism of AS.