Effects Of MiR-370-3p On The Proliferation Of Human Articular Chondrocytes By Regulating The JAK2-STAT5B Axis

Objective:To study non-coding single chain RNA(micro RNA,miRNA)miR-370-3p whether regulate the proliferation of human articular chondrocytes(HC-a)by JAK2(Janus kinase2)/ STAT5B(signal transduction and activator of transcription 5B)signaling pathway,and explore its potential mechanism of action.Method:(1)miR-370-3p over-expression/low-expression HC-a were constructed by liposome transfection,There were 5 groups: miR-370-3p mimic group(miR-370-3p overexpression group),miR-370-3p inhibitor group(miR-370-3p low-expression group),m NC(mimic normol control group),i NC(inhibitor normol control group),normal control group,(2)miRNA Quantitative real-time polymerase Chain Reaction was used to detect the relative expression of miR-370-3p in each group.(3)The relative mRNA expression of JAK2、STAT5B in each group was measured by qRT-PCR(Quantitative real-time polymerase Chain Reaction).(4)The expression level of JAK2、STAT5B protein in each group was detected western blot experiment.(5)MTT colorimetry was used to detect the relative proliferation of HC-a cells in each group.(6)The binding of miR-370-3p and JAK2、STAT5B was detected by dual luciferase reporter assay system experiment.Results :miRNA Quantitative real-time polymerase Chain Reaction experiment suggest that compared with the normal control group,The expression of miR-370-3p in mimic group was upregulated(P<0.05),The miR-370-3P expression in inhibitor group was downregulated(P<0.05),A model of miR-370-3p overexpression/low expression chondrocytes was successfully constructed;qRT-PCR results suggest that compared with the normal control group,The relative expression of JAK2 and STAT5 B in mimic group was downregulated and was upregulated in the inhibitor group(P<0.05).There was no significant difference between the empty vector control group and the normal control group.Western blot experiment showed significant downregulation of JAK2 and STAT5 B in the mimic group at the protein level,which was upregulated in the inhibitor group.MTT experiments suggest that miR-370-3p overexpression reduces HC-a survival rate(P<0.05),suggesting that miR-370-3p over-expression can inhibit human articular chondrocyte(HC-a)proliferation.The results of the Dual luciferase reporter assay system experiment showed that,following miR-370-3p and JAK2-WT(wild type),STAT5B-WT(wild type)plasmids were cotransfected,the fluorescence expression was significantly down-regulated compared with the control group,,miR-370-3p can directly combine with the JAK2-3’UTR、STAT5B-3’ UTR to regulate the signal pathway.Conclusion: miR-370-3P can inhibit the proliferation of human articular chondrocytes(HC-α)by directly targeting the regulation of JAK2/STAT5 B signaling pathway.