Predicted Mmu-miR-30α Regulation Of Glucocorticoid Receptor And In The Expression Of Mouse Podocytes

PartⅠBioinformatics prediction GR mRNA is target gene of mmu-miR-30aObjective:To probe whether mmu-mir-30a target gene is gluco-corticoid receptor (Glucocorticoid Receptor, GR) mRNA.Methods:Using miRBase (http://www.mirbase.org) to get miR-30a gene sequence; using Pubmed (http://www.ncbi.nlm.nih.gov) to get gene NR3C1 sequence; using miRanda, TargetScan, PicTar bioinformatics software for mmu-miR-30a target gene search and prediction.Results:All of miRanda, TargetScan and PicTar predicted that mmu-mir-30a target gene is the glucocorticoid receptor (Glucocorticoid Receptor, GR) mRNA.Conclusion:GR mRNA might be mmu-miR-30a target genePartⅡThe change of GRa and mmu-miR-30a expression in podocyteObjective:To investigate the expression of GRa and mmu-miR-30a in conditional immortalized mouse podocyte clone 5 (MPC5).Method:Conditionally immortalized murine podocyte cell line was cultured under permissive condition at 33℃.After passage the cells were shifted to non-permissive eondition at 37℃and cultured for 14 days.The MPC5 were divided into two groups,the expression of nephrin, Synap-topodin and GRa in undifferentiated MPC5 and differentiated MPC5 was detected by RT-PCR,Western-blot and Immunofluorescence.The expression of mmu-miR-30a in undifferentiated MPC5 and differentiated MPC5 was detected by qRT-PCR.Results:MPC5 cells can be cultured in two different phenotypes.MPC5 cells grew under permissive conditions (at 33℃with 100 U/mlγ-interferon).These cells proliferate and maintain an epithelial phenotype with cobblestone-like morphology.MPC5 cells grew under nonpermissive conditions (10-14 days at 37℃,noγ-interferon).In contrast,culturing under nonpermissive conditions stopped proliferation and induced conversion into arborized cells.The expression of nephrin was detected in 33℃MPC5 group and 37℃MPC5 group by RT-PCR,Western-blot and Immunofluorescence. Compared to 33℃MPC5 group,nephrin expression level in 37℃MPC5 group increased significantly (P<0.01); The expression of Synaptopodin was obviously detected in 37℃MPC5 group but nearly no expression in 33℃MPC5 group by RT-PCR,Western-blot and Immunofluorescence. (P<0.01);The expression of GRa was detected in 33℃MPC5 group and 37℃MPC5 group by RT-PCR,Western-blot and Immunofluorescence. Compared to 33℃MPC5 group,GRa expression level in 37℃MPC5 group increased significantly (P<0.01); The expression of mmu-miR-30a was detected in 33℃MPC5 group and 37℃MPC5 group by qRT-PCR. Compared to 33℃MPC5 group, mmu-miR-30a expression level in 37℃MPC5 group increased significantly (P<0.01).Conclusions:Mmu-miR-30a expression level was changed when MPC5 differentiated...