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MC3T3-E1 cells were divided into Blank, miR-135b mimics, miR-135b inhibitors, AG490, and miR-135b inhibitors + AG490 groups. Cell viability was determined by MTT, alkaline phosphatase (ALP) activity by the corresponding kit, and mineralization by alizarin red staining. Furthermore, miR-135b, osteoblast-specific genes, and JAK2/STAT3 were detected through quantitative real-time polymerase chain reaction and Western blotting. MiR-135b downregulation was identified with increased JAK2 during osteoblast differentiation. JAK2 was confirmed as a target gene of miR-135b by dual-luciferase reporter assay. MC3T3-E1 cells in both miR-135b mimics and AG490 groups manifested decrease in cell viability, ALP activity, and mineralized nodes, as well as reductions in osteoblast-specific genes and proteins of JAK2, p-JAK2, and p-STAT3, but increase in cell apoptosis. However, opposite changes of the above factors were shown in cells from miR-135b inhibitors group. Notably, AG490 could reverse promotion effects of miR-135b inhibitors on osteoblast differentiation. Inhibiting miR-135b could activate the JAK2/STAT3 signaling pathway, thereby improving the cell viability and promoting the osteoblast differentiation.
Pubmed ID: 32319222
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Database of microRNA target predictions and expression profiles. Target predictions are based on a development of the miRanda algorithm which incorporates current biological knowledge on target rules and on the use of an up-to-date compendium of mammalian microRNAs. MicroRNA expression profiles are derived from a comprehensive sequencing project of a large set of mammalian tissues and cell lines of normal and disease origin. This website enables users to explore: * The set of genes that are potentially regulated by a particular microRNA. * The implied cooperativity of multiple microRNAs on a particular mRNA. * MicroRNA expression profiles in various mammalian tissues. The web resource provides users with functional information about the growing number of microRNAs and their interaction with target genes in many species and facilitates novel discoveries in microRNA gene regulation. The microRNA Target Detection Software, miRanda, is an algorithm for finding genomic targets for microRNAs. This algorithm has been written in C and is available as an open-source method under the GPL.
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