Research Paper Volume 12, Issue 21 pp 22291—22312

LncRNA PVT1 accelerates malignant phenotypes of bladder cancer cells by modulating miR-194-5p/BCLAF1 axis as a ceRNA

Mingwei Chen1,2, *, , Rongyuan Zhang1, *, , Le Lu1, , Jian Du1, , Chunyang Chen1, , Keke Ding1, , Xuedong Wei1, , Guangbo Zhang1,3, , Yuhua Huang1, , Jianquan Hou1, ,

  • 1 Department of Urology, The First Affiliated Hospital of Soochow University, Suzhou 215006, Jiangsu Province, China
  • 2 Department of Urology, The Fourth Affiliated Hospital, Zhejiang University School of Medicine, Yiwu 322000, Zhejiang Province, China
  • 3 Jiangsu Institute of Clinical Immunology, The First Affiliated Hospital of Soochow University, Jiangsu Key Laboratory of Clinical Immunology, Soochow University, Jiangsu Key Laboratory of Gastrointestinal Tumor Immunology, Suzhou 215006, Jiangsu Province, China
* Equal contribution

Received: April 10, 2020       Accepted: October 8, 2020       Published: November 16, 2020      

https://doi.org/10.18632/aging.202203
How to Cite
This article has been corrected. See Correction. Aging (Albany NY). 2021; 13:4731-4733. https://doi.org/10.18632/aging.202683  PMID: 33587046

Copyright: © 2020 Chen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Background: Numerous studies proved that long non-coding RNA (lncRNA) is involved in the progression of multifarious diseases, especially in some carcinomas. As a potential tumor biomarker, plasmacytoma variant translocation 1 gene (PVT1) is involved in the development and progression of multifarious cancers. Nevertheless, the intrinsic and concrete molecular mechanism of PVT1 in bladder cancer still remained unclear, which is also the dilemma faced in many non-coding RNA studies.

Results: Our research revealed that PVT1 was significantly higher expression in bladder carcinoma specimens and cell lines. Further experiments indicated that knockdown or overexpression of PVT1 restrained or promoted the malignant phenotype and WNT/β-catenin signaling in bladder cancer cells. Meanwhile miR-194-5p was in contrast and miR-194-5p could partially reverse the function of PVT1 in malignant bladder tumor cells. As a microRNA sponge, PVT1 actively promotes the expression of b-cells lymphoma-2-associated transcription factor 1 (BCLAF1) to sponge miR-194-5p and subsequently increases malignant phenotypes of bladder cancer cells. Therefore, it performs a carcinogenic effect and miR-194-5p as the opposite function, and serves as an antioncogene in the bladder carcinomas pathogenesis.

Conclusion: PVT1-miR-194-5p-BCLAF1 axis is involved in the malignant progression and development of bladder carcinomas. Experiments revealed that PVT1 has a significant regulatory effect on bladder cancer (BC) and can be used as a clinical diagnostic marker and a therapeutic molecular marker for patients suffering from BC.

Methods: In urothelial bladder carcinoma specimens and cell lines, the relative expression levels of PVT1 and miR-194-5p were detected by quantitative reverse transcription PCR (RT-qPCR). Through experiments such as loss-function and over-expression, the biological effects of PVT1 and miR-194-5p on the proliferation, migration, apoptosis and tumorigenicity were explored in bladder cancer cells. Co-immunoprecipitation, proteomics experiments, dual luciferase reporter gene analysis, western blot and other methods were adopted to investigate the PVT1 potential mechanism in bladder carcinomas.

Abbreviations

lncRNAs: Long non-coding RNAs; PVT1: plasmacytoma variant translocation 1 gene; qPCR: Quantitative polymerase chain reaction; TNM: Tumor Node Metastasis; BCLAF1: b-cell lymphoma-2-associated transcription factor 1; ceRNA: competing endogenous RNA; BC: bladder cancer/carcinoma; miRNAs: MicroRNAs; shRNAs: Short hairpin RNAs; shNC: Short hairpin negative control; siRNA: Small interfering RNA; co-IP/IP: co-immunoprecipitation; EdU: Ethynyl-2-deoxyuridine; CCK-8: Cell Counting Kit-8; EdU: Ethynyl-2-deoxyuridine; si-NC: Negative Control siRNA; BC: Bladder cancer; FBS: Fetal bovine serum; CCK8: Cell Counting Kit-8; SDS-PAGE: SDS-polyacrylamide gel electrophoresis; PVDF: Polyvinylidene Fluoride; IDVs: Integrated density values; ANOVA: Analysis of Variance; LV-shRNA: Lentivirus shRNA.