MiR 3180-5p promotes proliferation in human bladder smooth muscle cell by targeting PODN under hydrodynamic pressure.
Human bladder smooth muscle cells (HBSMCs) were subjected to pressure cycles of up to 200 cm H2O to a pressure of 0 cm H2O for 24 hours. 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 (p = 0.004 and p = 0.041, respectively). Silencing of PODN via miR-3180-5p overexpression revealed a significant promotion of cell proliferation increased in the CCK-8 experiment, p = 0.00077). This cell proliferation was accompanied by an increase in cdk2 expression (p = 0.00193) and a decrease in p21 expression (p = 0.0095). The percentage of cells in (S + G2/M) improved after transfection (p = 0.002). It was apparent that HP upregulates miR-3180-5p, which inhibits the expression of PODN and promotes HBSMC proliferation via the cdk2 signaling pathway.
Department of Urology, West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.|Institute of Urology (Laboratory of Reconstructive Urology), West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.
Yi
Y
Sun
Department of Urology, West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.|Institute of Urology (Laboratory of Reconstructive Urology), West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.
De-Yi
DY
Luo
Department of Urology, West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.|Institute of Urology (Laboratory of Reconstructive Urology), West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.
Yu-Chun
YC
Zhu
Department of Urology, West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.|Institute of Urology (Laboratory of Reconstructive Urology), West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.
Liang
L
Zhou
Department of Urology, West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.|Institute of Urology (Laboratory of Reconstructive Urology), West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.
Tong-Xin
TX
Yang
Department of Urology, West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.|Institute of Urology (Laboratory of Reconstructive Urology), West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.
Cai
C
Tang
Department of Urology, West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.|Institute of Urology (Laboratory of Reconstructive Urology), West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.
Hong
H
Shen
Department of Urology, West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.|Institute of Urology (Laboratory of Reconstructive Urology), West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China.
Kun-Jie
KJ
Wang
D002460:Cell Line; D049109:Cell Proliferation; D051357:Cyclin-Dependent Kinase 2 / Q000378:metabolism; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D006874:Hydrostatic Pressure; D036341:Intercellular Signaling Peptides and Proteins; D020935:MAP Kinase Signaling System; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism*; D001743:Urinary Bladder / Q000166:cytology / Q000378:metabolism*
D006023:Glycoproteins; D036341:Intercellular Signaling Peptides and Proteins; D035683:MicroRNAs; C477527:PODN protein, human; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2