miRNA在人骨髓来源间充质干细胞成骨诱导分化过程中的表达

miRNA Expression during Osteo-differentiation of Human Bone Marrow-derived Mesenchymal Stem Cells

观察miRNAs在人骨髓来源间充质干细胞成骨诱导分化过程中的表达情况。以从骨髓中分离培养的MSCs及成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。分离培养出的MSCs经成骨诱导培养14 d后,已具有成骨细胞特性;经芯片检测并SAM分析,成骨诱导培养的细胞较MSCs高表达的miRNAs有7个:hsa-miR-363*、hsa-miR-483、hsa-miR-590、hsa-miR-130a、hsa-miR-15b、hsa-miR-30c、hsa-miR-663;成骨诱导培养的细胞较MSCs低表达的miRNAs有6个:hsa-miR-192、hsa-miR-210、hsa-miR-128a、hsa-miR-224、hsa-miR-106a、hsa-miR-494。利用TargetScan预测其靶基因,并行生物信息学分析,其中hsa-miR-130a、hsa-miR-15b和hsa-miR-30c的预测靶基因多为维持干细胞特性的基因;而hsa-miR-106a和hsa-miR-494的预测靶基因多为参与骨形成及成骨分化的基因。为证明miRNAs参与调控MSCs的成骨分化过程提供了直接证据和研究基础。

To identify miRNA expression during osteo-differentiation of human bone marrow-derived mesenchy- mal stem cells, MSCs were isolated and cultured by using density centrifugation from bone marrow, and induced to differentiate into osteoblasts. Using miRNA microarrays single-channel fluorescence chip from CapitalBio Corpora-tion （Beijing, China） , samples of MSCs and osteoblasts were assayed to determine expression levels for miRNAs. Raw data were analyzed using the Significance Analysis of Microarrays （ SAM, version 2.1 ） software to determine miRNAs expressed in MSCs and osteoblasts. MSCs were isolated from bone marrow and induced to differentiate into osteoblasts in vitro culture, miRNA expression of MSCs and osteoblasts were investigated using microarrays. Then 7 miRNAs overexpressed in osteoblasts and （underexpressed in MSCs） ： hsa-miR-363 ＊ ,hsa-miR-483 ,hsa-miR-590, hsa-miR-130a,hsa-miR-15b, hsa-miR-30c, hsa-miR-663 ; and 6 miRNAs underexpressed in MSCs （ overexpressed in MSCs） ： hsa-miR-192,hsa-miR-210,hsa-miR-128a,hsa-miR-224,hsa-miR-106a,hsa-miR-494. To predict their target genes by using bioinformatics software, and these target genes were for osteogenic differentiation and sternness maintaining. These finding indicate that miRNAs are likely important regulators for osteo-differentiation of MSCs.