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目的:研究miRNAs在人骨髓来源间充质干细胞软骨诱导分化过程中的表达情况。方法:以从骨髓中分离培养的MSCs及软骨诱导培养后的细胞为实验对象,利用基因芯片检测miRNAs的表达情况,由SAM分析得到MSCs较其诱导培养细胞中差异表达的miRNAs,再进行生物信息学分析。结果:①分离培养出的MSCs经软骨诱导培养21天后,已具有软骨细胞特性,经芯片检测并SAM分析,软骨诱导培养的细胞较MSCs高表达的miRNAs有6个:hsa-miR-572、hsa-miR-130b、hsa-miR-193b、hsa-miR-28、hsa-miR-152、hsa-miR-560;软骨诱导培养的细胞较MSCs低表达的miRNAs有2个:hsa-miR-424、hsa-miR-122a。②利用TargetScan预测其靶基因,并行生物信息学分析,其中hsa-miR-130b、hsa-miR-193b、hsa-miR-152及hsa-miR-424的预测靶基因中多为参与细胞分化、骨形成、软骨形成及干细胞表型相关的基因。结论:hsa-miR-130b、hsa-miR-193b、hsa-miR-152和hsa-miR-424等对人骨髓来源间充质干细胞的软骨分化起着重要调控作用。
OBJECTIVE: To study the expression of miRNAs in the process of chondrocyte differentiation induced by human bone marrow mesenchymal stem cells. Methods: MSCs isolated from bone marrow and cartilage were used to induce the cultured cells. The miRNAs were detected by microarray. Microarrays differentially expressed in cultured MSCs were analyzed by SAM, and bioinformatics Analysis. Results: (1) Chondrocytes were isolated from chondrocytes induced by chondrocytes cultured for 21 days. Chips were detected by SAM and analyzed by SAM. There were 6 miRNAs highly expressed in cartilage-induced cells compared with MSCs: hsa-miR-572, hsa miR-130b, hsa-miR-193b, hsa-miR-28, hsa-miR-152 and hsa-miR- hsa-miR-122a. (2) TargetScan was used to predict the target genes, and bioinformatics analysis was performed in parallel. Most of the predicted target genes of hsa-miR-130b, hsa-miR-193b, hsa-miR-152 and hsa-miR-424 were involved in cell differentiation, Formation, chondrogenesis and stem cell phenotype related genes. Conclusion: hsa-miR-130b, hsa-miR-193b, hsa-miR-152 and hsa-miR-424 play an important role in the regulation of cartilage differentiation of human bone marrow-derived mesenchymal stem cells.