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Objective To explore the effect of long noncoding RNA-ATB (LncRNA-ATB) on phenotypic transition and proliferation of human peritoneal mesothelial cells (HPMCs) induced by high glucose. Methods HPMCs used in experiment were divided into three groups: control group, mannitol group and hypertonic glucose group. HPMCs in control group received no treatment, and in hypertonic glucose group and mannitol group were treated with 50mmol/L D-glucose and isotonic mannitol for 72 hours, respectively. Real-time PCR was employed to detect the mRNA expression of LncRNA-ATB, E-cadherin, α-smooth muscle actin (α-SMA), connective tissue growth factor (CTGF), Cyclin D1, cyclin dependent kinase inhibitor 4 (CDK4), protein 27 (p27) and proliferating cell nuclear antigen (PCNA). Western blotting was performed to detect the proteins expression of E-cadherin, α-SMA, CTGF, Cyclin D1, CDK4, p27 and PCNA, and flow cytometry was used to test the cell cycle. Lentivirus artifice was used to up- or down-regulate the expression of LncRNA-ATB in untreated HPMCs. Real-time PCR was employed to detect the mRNA expression of E-cadherin, α-SMA and CTGF, Western blotting was performed to detect the proteins expression of E-cadherin, α-SMA and CTGF, and flow cytometry was used to test the cell cycle. Results It is revealed by Real-time PCR, Western blotting and flow cytometry that the expressions increased of LncRNA-ATB, α-SMA, CTGF, Cyclin D1, CDK4 and PCNA induced by hypertonic glucose, and decreased of E-cadherin and p27 (P |
