软骨细胞SHOX基因过／低表达的差异miRNA筛选与验证  

Screening and verification of differential miRNAs in chondrocytes with overexpression and low expression of SHOX

作　　者：许珂[1] 徐雪姣 宋萃[1] 朱岷[1] 

机构地区：[1]重庆医科大学附属儿童医院内分泌科儿童发育疾病研究教育部重点实验室儿童发育重大疾病国家国际科技合作基地儿科学重庆市重点实验室,重庆400014

出　　处：《第三军医大学学报》2018年第3期211-215,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81170723)

摘　　要：目的建立软骨细胞(HC-α)SHOX基因过/低表达模型,筛选与未处理组差异表达miRNA,并予以验证。方法通过慢病毒过/低表达SHOX基因,用miRNA芯片筛选差异表达的miRNA。结果 SHOX过表达组与对照组有425条差异表达的miRNA;SHOX低表达组与对照组有379条差异表达的miRNA;按照表达水平差异相反的标准筛选差异miRNA,其中6条miRNA在处理组中表达水平差异相反。实时荧光定量PCR对6条miRNA(miR-126、miR-192、miR-370、miR-432、miR-3162、miR-4745)进行验证,miR-126在过表达组明显上调(P<0.05),在低表达组明显下调(P<0.05)。结论 miR-126参与SHOX介导的软骨细胞凋亡过程。Objective To establish 2 cell models with overexpression and low expression of short stature homeo-box-containing gene （SHOX） in chondrocytes and screen the differentially expressed miRNAs in the cells. Methods Lentiviral vectors were used to establish a cell model of SHOX overexpression and a cell model of SHOX gene interference in HC-α cells. Microarray chip technique was used to screen the differentially expressed miRNAs in the 2 cell models compared with HC-α cells without genetic modification （NC group）. ResultsCompared with the NC group, 425 and 379 differentially expressed miRNAs were identified in HCα cells with SHOX overexpression and in cells with SHOX interference, respectively. Six miRNAs were found to have opposite changes between the 2 cell models, and verification of the 6 miRNAs （miR-126, miR-192, miR-370, miR-432, miR-3162, and miR-4745） with real-time PCR confirmed that miR-126 was significantly up-regulated （P〈0.05） in cells with SHOX overexpression and was significantly down-regulated （P〈0.05） in cells with SHOX interference. Conclusion miR-126 is involved in SHOX-mediated chondrocyte apoptosis.

关 键 词：SHOX 软骨细胞 MIRNA芯片 

分 类 号：R322.71[医药卫生—人体解剖和组织胚胎学] R394.33[医药卫生—基础医学]