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Effects of miR-29a on epithelial to mesenchymal transition in human peritoneal mesothelial cells
2018, 17 (10):
694-698.
doi: 10.3969/j.issn.1671-4091.2018.10.010
【Abstract】Objective To investigate the effect of miR-29a on high glucose induced epithelial to mesenchymal transition (EMT) in human peritoneal mesothelial cells (HPMCs). Methods HPMCs were treated with high glucose to create a EMT model of HPMCs. Real-time PCR was used to quantify the expression of miR-29a and EMT makers including E-cadherin, α-SMA and fibronectin (FN) in HPMCs stimulated with various concentration of glucose for various period as well as in control cells. miR- 29a inhibitor was used to downregulate miR-29a expression in HPMCs, and the expression of miR-29a and EMT markers in these cells were then measured by real-time PCR. Results High glucose stimulated HPMCs for various concentration and period promoted the expression of α-SMA (1.5% glucose: 1.970±0.153 vs. 1, F=7.692, P=0.008; 2.5% glucose: 3.291±0.265 vs. 1, F=4.479, P<0.001; 4.25% glucose: 4.301±0.346 vs. 1, F=5.496, P<0.001; high glucose for 6h: 2.161±0.202 vs. 1, F=6.563, P=0.001; high glucose for 12h: 2.820±0.285 vs. 1, F=5.111, P<0.001; high glucose for 24h: 3.291±0.265 vs. 1, F=4.479, P<0.001; high glucose for 48h: 3.980±0.407 vs. 1, F=9.3769, P=0.006) and FN (1.5% glucose: 1.630 ± 0.157 vs. 1, F=7.092, P=0.002; 2.5% glucose: 2.910 ± 0.199 vs. 1, F=4.000, P<0.001; 4.25% glucose: 3.601±0.301 vs. 1, F=5.575, P<0.001; high glucose for 6h:1.761±0.172 vs. 1, F=7.144, P=0.002; high glucose for 12h: 2.390±0.170 vs. 1, F=4.499, P<0.001; high glucose for 24h: 2.910±0.199 vs. 1, F=4.000, P<0.001; high glucose for 48h: 3.601±0.300 vs. 1, F=4.570, P<0.001), but inhibited the expression of E-cadherin (1.5% glucose: 0.693±0.065 vs. 1, F=4.665, P=0.001; 2.5% glucose: 0.452±0.045 vs. 1, F=4.994, P<0.001; 4.25% glucose: 0.302±0.030 vs. 1, F=4.000, P<0.001; high glucose for 6h: 0.802±0.084 vs. 1, F=4.000, P=0.012; high glucose for 12h: 0.630±0.070 vs. 1, F=7.030, P=0.001; high glucose for 24h: 0.452±0.045 vs. 1, F=4.994, P<0.001; high glucose for 48h: 0.290±0.030 vs. 1, F=4.000, P<0.001), suggesting that EMT occurred in HPMCs stimulated with high glucose. Moreover, miR-29a expression increased in HMPCs along with the increase of glucose concentration and stimulation period (1.5% glucose: 1.452 ± 0.147 vs. 1, F=13.411, P=0.001; 2.5% glucose: 3.120 ± 0.320 vs. 1, F=10.372, P<0.001; 4.25% glucose: 4.130±0.368 vs. 1, F=12.497, P<0.001; high glucose for 6h: 1.330±0.114 vs. 1, F=5.788, P=0.001; high glucose for 12h: 2.310±0.173 vs. 1, F=5.546, P<0.001; high glucose for 24h: 3.100±0.236 vs. 1, F=10.372, P<0.001; high glucose for 48h: 3.310 ± 0.211 vs. 1, F=4.929, P<0.001). When HPMCs were transfected with the miR-29a inhibitor to inhibit the expression of miR-29a, the cells showed decreased expression of α-SMA and FN and increased expression of E- cadherin, indicating that the EMT induced by high glucose was inhibited in these cells (α-SMA: 1.871±0.206 vs. 3.291±0.265, F=0.104, P=0.002; FN: 1.782±0.156 vs. 2.910±0.199, F=0.091, P=0.002; E-cadherin: 0.873±0.085 vs. 0.452±0.045, F=0.760, P=0.002). Conclusion High glucose induced EMT in dose dependent and time dependent manner. In addition, there was a positive correlation between miR-29a and EMT; EMT induced by high glucose was attenuated in HPMCs in which miR-29a was downregulated.
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