应用基因表达芯片筛选大骨节病软骨差异表达的基因和基因通路  

Identification of differently expressed genes and pathways in cartilage of Kashin-Beck disease using microarray

作　　者：张峰[1] 王伟卓 郭雄[1] 武世勋[1] 王立新[1] 

机构地区：[1]西安交通大学医学院环境与疾病相关基因教育部重点实验室及卫生部微量元素与地方病重点实验室,710061 [2]第二附属医院骨科

出　　处：《中华地方病学杂志》2013年第2期159-163,共5页Chinese Journal of Endemiology

基　　金：国家自然科学基金（30972556,81102086）;陕西省“13115”科技创新工程重大科技专项（2009ZDKG-79）;高等学校博士学科点专项科研基金（20110201120057）

摘　　要：【摘要】目的筛选大骨节病（Kashin—Beckdisease，KBD）和正常关节软骨的差异表达基因和基因通路，探讨KBD关节软骨损伤的分子机制。方法KBD组和对照组关节软骨样本各9份，提取总RNA，反转录为eDNA。采用Cy3、Cy5分别对KBD组、对照组进行荧光标记。采用Agilent全基因组表达芯片，比较KBD组与对照组关节软骨的表达谱差异，利用单基因及基因通路表达分析，筛选有统计学意义的差异表达的基因和基因通路。结果①KBD组的关节软骨中29个基因的表达水平显著上调（平均表达率=6．68±1．98，P均〈0．05），功能涉及细胞凋亡、代谢、细胞外基质、细胞骨架与运动等。KBD患者关节软骨中的细胞外基质相关的FBLNl基因表达水平显著下调（表达率=0．14±0．06，P〈0．05）。@KBD组关节软骨中6个凋亡和5个缺氧相关的基因通路的表达水平均高于对照组（P均〈0．05）。结论细胞凋亡和缺氧相关的基因及基因通路在KBD患者与对照关节软骨中的表达水平存在差异，提示缺氧可能在KBD关节软骨细胞凋亡中发挥一定作用，缺氧与KBD关节软骨损伤的关系有待进一步研究。Objective To identify differently expressed genes and pathways between Kashin-Beck disease （KBD） cartilage and healthy cartilage, and to explore the mechanism of articular Cartilage lesions of KBD. Methods Cartilage specimens were collected from 9 patients with KBD and 9 healthy controls. Total RNA was extracted from cartilage specimens, and transcribed into cDNA. KBD and control groups were labeled by Cy3 and Cy5, respectively. Agilent genome-wide microarray was applied to compare the expression profile of KBD cartilage and healthy cartilage. The microarray data was analyzed by single gene and pathway expression analysis to identify differently expressed genes and pathways between KBD and healthy controls. Results ①Tweenty nine genes were significantly up-regnlated in KBD group （averaged ratio = 6.68 ± 1.98, P 〈 0.05）, mainly involved in apoptosis, metabolism, extracellular matrix, cytoskeleton and cell movement. Additionally, extracellular matrix-related FBLN1 gene was down-regulated in KBD group（ratio = 0.14 ± 0.06, P 〈 0.05）.②Five apoptosis and 6 hypoxia-related pathways presented higher expression levels in KBD compared to healthy controls（all P〈 0.05）. Conclusions We find significant expression differences of apoptosis and hypoxia-related genes and pathways between KBD cartilages and healthy cartilages, suggesting that hypoxia might contribute to chondrocytes apoptosis of KBD. Further studiesmay be needed to investigate the relationship between hypoxia and articular cartilage lesions of KBD.

关 键 词：大骨节病 缺氧 细胞凋亡 基因 寡核苷酸序列分析 

分 类 号：R684.1[医药卫生—骨科学]