Influence Of Mesenchymal Stem Cells Of Bone Marrow Of MiRNAs

MiRNA is a kind of short, non-coding,single-stranded small RNA.It plays an important role through post-transcriptional regulation of gene expression. Several studies showed that miRNA have important functions of regulation in cell proliferation, differentiation, apoptosis, secretion, migration, malignant transformation and other cellular functions. Bone marrow stem cells(BMSC) are a class of pluripotent stem cells, they can differentiate into a variety of mesodermal or non-mesodermal cells. Many studies have shown that the differentiat abnormalities of BMSC took part in osteonecrosis diseases like femoral head necrosis, osteoporosis and some other diseases. MiRNA regulates the direction of differentiation of bone marrow stem cells by regulate a large amount of genes by a post transcriptional protocol. In this study, we carry out a functional verification on the basis of our previously screened miRNA in order to clarify their influence on the direction of bone marrow stem cells.Objects1. Recovery, culture, passage of BMSC, and screen miRNA using bioinformatics method on the basis of our previous studies;2. Custom agomir and antagomir to verify microRNA function by transfection. After changing the corresponding miRNA levels of bone marrow stem cells, test the differential direction of mesenchymal stem cell by Oil Red O, alizarin red, and type Ⅱ collagen staining. Thus screened the miRNA which has infect on bone marrow stem cells;3. Using whole-genome microarray to study the expression of target genes regulated by miRNA.MethodsUsing bone marrow stem cells derived by ScienCell, after recovery, culture and passage, we obtained P3-P8of bone marrow stem cells. Using agomir and antagomir to transfect bone marrow stem cells. After96hours of transfection,we test the differential direction of mesenchymal stem cell by Oil Red0, alizarin red, and type Ⅱ collagen staining. Thus we observed the osteogenic, adipogenic, chondrogenic function of transfectod bone marrow stem cells.After that,we extract RNA for microarray.Results1. After recovery, training, passage of bone marrow stem cells, we obtained P3-P8generation of bone marrow stem cells; according to the previous test results of bioinformatics, we screened8miRNAs may impact the differentiation;2. According to the transfeetion and staining of oil red O, alizarin red, type Ⅱ collagen, Hsa-miR-1303can regulate the function of bone marrow stem cell differentiation;3. Using whole-genome microarray we obtained the gene expression of transfected bone marrow stem cell,so we arriving the target gene of Hsa-miR-1303:ANKH.Conclusion:In these8miRNAs seleced by Bioinformatics, hsa-mir-1303can regulate the differentiation of bone marrow stem cells. Using whole-genome microarray we identify the target gene of has-mir-1303:ANKH. According to the functional enrichment analysis, hsa-miR-1303get the function of the regulation of osteoclast differentiation.