[No authors listed]
Human bladder smooth muscle cells (HBSMCs) were subjected to pressure cycles of up to 200âcm H2O to a pressure of 0âcm H2O for 24âhours. The total RNA extracted from each group was subjected to microarray analysis. miR-3180-5p emerged as the most overexpressed of all the differentially expressed microRNAs, and this finding was validated by PCR. We then used CCK-8 to quantify cell proliferation after liposome-mediated transfection. Subsequently, we investigated the change in PODN and its downstream signaling proteins, including cyclin-dependent kinase 2 (cdk2) and p21. In addition, flow cytometry was performed to quantify cell-cycle distribution. The results show that miR-3180-5p, the microRNA that was most overexpressed in response to HP, reduced the expression of PODN and podocan (pâ=â0.004 and pâ=â0.041, respectively). Silencing of PODN via miR-3180-5p overexpression revealed a significant promotion of cell proliferation increased in the CCK-8 experiment, pâ=â0.00077). This cell proliferation was accompanied by an increase in cdk2 expression (pâ=â0.00193) and a decrease in p21 expression (pâ=â0.0095). The percentage of cells in (Sâ+âG2/M) improved after transfection (pâ=â0.002). It was apparent that HP upregulates miR-3180-5p, which inhibits the expression of PODN and promotes HBSMC proliferation via the cdk2 signaling pathway.
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