退变性脊柱侧凸髓核组织中microRNA筛选及靶基因定位

Screening and target gene mapping of differentially expressed microRNAs in the degenerative intervertebral disc of degenerative scoliosis patients

背景:microRNAs(miRNAs)在多种疾病中扮演重要的角色,对退变性脊柱侧凸患者mi RNAs表达谱的分析,有助于揭示其发病机制。目的:比较退变性脊柱侧凸患者与正常对照组椎间盘组织中mi RNAs表达谱的差异,并探究其在退变性脊柱侧凸发病机制中的作用。方法:对获取的48例退变性脊柱侧凸患者(男36例,女12例;58-69岁)术中切除的椎间盘组织髓核组织和36例腰椎骨折患者正常髓核组织依次进行髓核细胞分离、培养、染色鉴定、提取标本总RNA;采用microRNA微阵列基因表达实验和分析技术筛选差异表达的mi RNAs并采用实时q PCR技术对其中高表达者进行验证;综合MicroCosmv5、Target Scan 5.1和microRNA.org等3个数据库的靶基因信息,取交集分析预测靶基因,并分析与差异靶基因功能显著相关的生物信号通路;定量PCR方法验证筛选结果。结果与结论:19条mi RNA表达存在差异。经验证后,退变性脊柱侧凸患者的椎间盘组织中mi R-98呈显著高表达,倍值变化为6.368;预测靶基因为白细胞介素10,此靶基因位于JAK-STAT信号通路的上游,参与其信号传导。miR-98在退变性脊柱侧凸患者的椎间盘中呈高表达,对应的靶基因为白细胞介素10。提示该靶基因启动JAK-STAT信号通路在退变性脊柱侧凸的发病机制中可能发挥至关重要的作用。

BACKGROUND：MicroRNAs （miRNAs） play an important role in many diseases. To analyze the miRNA expression profile in degenerative scoliosis patients is helpful for classifying its pathogenesis. OBJECTIVE： To compare miRNAs expression profile in the intervertebral disc tissue between degenerative scoliosis patients and healthy controls, and to investigate its role in the pathogenesis of degenerative scoliosis. METHODS：Degenerative nucleus pulposus tissues from 48 patients with degenerative scoliosis （male 36, female 12; 58-69 years old） and normal nucleus pulposus tissues from 36 patients with lumbar fractures were harvested to isolate, culture and identify nucleus pulposus cels folowed by total RNA extract. Differentialy expressed miRNAs were screened by microRNA microarray analysis and validated by real-Time qPCR. Target genes of highly expressed microRNAs were predicted by analyzing information from MicroCosm v5, TargetScan 5.1 and microRNA.org databases. Biological signal pathways associated with the target genes were analyzed, and qPCR was used to validate the screening results. RESULTS AND CONCLUSION：Nineteen differentialy expressed miRNAs were identified. The miR-98 was highly expressed in degenerative nucleus pulposus tissue, and the fold change was 6.368. Predicted miR-98 target gene was interleukin-10, which was involved in JAK-STAT signaling pathway and located in upstream of this pathway. In degenerative nucleus pulposus cels of degenerative scoliosis patients, miR-98 was highly expressed, and the corresponding target gene was interleukin-10. These results indicate that JAK-STAT signaling pathway may play an important role in the pathogenesis of degenerative scoliosis.