Volume 29, Issue 3
DOI: 10.24205/03276716.2020.729
Regulatory Effects of Long-Chain Noncoding RNA PCGEM1 On Invasion and Metastasis of Bladder Urothelial Cell Carcinoma Cells Via the TGF Î’/Smad Signalling Pathway Running title: Lncrna And Bladder Urothelial Cell Carcinoma
Abstract
Background: Long-chain noncoding RNA (lncRNA) highly expressed in prostate cancer cells and acts as prostate cancer gene expression marker 1 (PCGEM1). However, the role of PCGEM1 in bladder urothelial cell carcinoma (BUCC) remains largely unknown. Objectives: Basic aim of current investigation is to analyse the regulatory effects of lncRNA PCGEM1 on the invasion and metastasis of BUCC cells via the TGF β/Smad pathway.
Materials and Methods: BUCC tissues and normal tissues over 2 cm away were collected from 128 BUCC patients who received surgery from March 2016 to May 2018. The expressions of lncRNA PCGEM1 in BUCC along with surrounding normal tissues as well as different BUCC cell lines were detected by qRT-PCR. LncRNA PCGEM1-silenced T24siPCGEM1 cells and negative control T24-siNC cells were constructed, using T24 cell line as blank control. Proliferation of T24 cell was examined by MTT and colony formation assays, and invasion and migration were detected by Transwell and scratch assays, respectively. The bioinformatics website starBase was used to predict miRNAs which complementarily bound PCGEM1, and miRNA-targeted binding genes were predicted through the www.microRNA.org website. The expressions of proteins in the TGF β2/Smad2 pathway were measured by Western blotting.
Results: qRT-PCR indicated about the expression of lncRNA PCGEM1 in BUCC tissues, which is higher than the surrounding normal tissues (P<0.05). LncRNA PCGEM1 had the highest expression in T24 cells. The optical density of the T24-siPCGEM1 group at 492 nm significantly decreased than blank control and T24-siNC groups. The numbers of colonies and cells penetrating Matrigel, together with the cell motility and wound healing rate all significantly decreased (P<0.05). LncRNA PCGEM1 complementarily bound miR-148a, and there was a targeted binding site between miR-148a and TGF β2. The expressions of TGF β2 and p-Smad2 in the T24-siPCGEM1 group were significantly lower but the expression of miR-148a was higher than those in blank and negative control groups (P<0.05).
Conclusions: LncRNA PCGEM1 is highly expressed in BUCC. High lncRNA PCGEM1 expression may promote the progression of BUCC by down-regulating miR-148a expression and enhancing the TGF β2/Smad2 signaling pathway.
Keywords
lncRNA; PCGEM1; miR-148a; bladder urothelial cell carcinoma; TGF β; Smad