Effect of down-regulation of plasmacytomas variant translocation gene 1 expression on proliferation of mesangial cells under high glucose

LI Ming; WU Wangli; PENG Yun; XIAO Yiping

doi:10.3969/j.issn.1002-266X.2017.14.003

Abstract：Objective To observe the effect of down-regulation of plasmacytomas variant translocation gene 1 (PVT1) in long non-coding RNA (lncRNA) on the proliferation of mesangial cells and the expression of extracellular matrix (ECM)-related proteins under high glucose;and the role of PVT1 in the pathogenesis of diabetic nephropathy.Methods We cultured human mesangial cells in vitro.Then;the cells were divided into normal control group;high glucose group;high glucose +control siRNA group;and high glucose +PVT1 siRNA group.The cells in the normal control group were cultured in 5.55 mmol/L DMEM medium;and cells in the other groups were cultured in 25 mmol/L DMEM medium.Cells in the high glucose +control siRNA group and high glucose +PVT1 siRNA group were transfected with control siRNA and PVT1 siRNA;respectively.After 0;24;48 and 72 h;the cell proliferation rate was measured by CCK-8.Flow cytometry was used to detect the proportion of cells in the G1 phase and S phase in each group.PVT1 mRNA expression in each group was detected by qRT-PCR.The expression of ECM protein was detected by Western blotting;including fibronectin (FN);transforming growth factor β1 (TGFβ1) and plasminogen activator inhibitor 1 (PAT-1).Results At each time point;the proliferation rate of cells in the high glucose group was higher than that in the normal control group;and the proliferation rate of high glucose +PVT1 siRNA was lower than that in the high glucose group (all P<0.05).Compared with the normal control group;the proportion of cells in the G1 phase of the high glucose group was decreased;and the proportion of cells in the S phase was increased;compared with the high glucose group;the the proportion of cells in the G1 phase in the high glucose + PVT1 siRNA group was increased;and the proportion of cells in the S phase was decreased (all P<0.05).The expression of PVT1 mRNA in the high glucose group and high glucose+control siRNA group was higher than that in the normal control group;and the expression of PVT1 mRNA in the high glucose+PVT1 group was lower than that in the high glucose group (all P<0.05).The expression of FN;TGF-β1and PAT-1 protein in the high glucose group;high glucose +control siRNA group and high glucose +PVT1 siRNA group was higher than that in the normal control group;the expression of FN;TGF-β1and PAT-1 protein in the high glucose+PVT1 group was lower than that in the high glucose group (all P<0.05).Conclusion Under the high glucose conditions;the expression of PVT1 in mesangial cells is significantly increased;and down-regulation of PVT1 expression could inhibit the proliferation of mesangial cells and inhibit the expression of ECM-related proteins.