MicroRNA‑16‑5p/BIMP1/NF‑κB axis regulates autophagy to exert a tumor‑suppressive effect on bladder cancer

Mol Med Rep. 2021 Aug;24(2):576. doi: 10.3892/mmr.2021.12215. Epub 2021 Jun 16.

Abstract

Bladder cancer (BC) is the second most common urological disease worldwide. Previous studies have reported that microRNA (miR)‑16‑5p is associated with the development of BC, but whether miR‑16‑5p regulates BC cell autophagy remains unknown. Thus, the aim of the present study was to investigate this issue. miR‑16‑5p expression in BC cells was assessed by reverse transcription‑quantitative PCR. Cell viability and apoptosis were detected via Cell Counting Kit‑8 and flow cytometry assays, respectively. For cell autophagy detection, autophagic flux was detected using a mCherry‑green fluorescent protein‑microtubule‑associated proteins 1A/1B light chain 3B (LC3) puncta formation assay, followed by determination of autophagy‑related protein markers. The targeting relationship between miR‑16‑5p and caspase recruitment domain family member 10 (BIMP1) was confirmed using a dual‑luciferase reporter assay, followed by detection of the BIMP1/NF‑κB signaling pathway. The results showed that miR‑16‑5p overexpression inhibited cell viability, whereas miR‑16‑5p knockdown promoted cell viability in BC. Furthermore, miR‑16‑5p overexpression induced autophagy, which was accompanied by increased autophagic flux and expression of the autophagy‑related proteins LC3‑II and beclin 1, as well as decreased p62 expression, whereas miR‑16‑5p silencing led to an inhibition of autophagy in BC cells. Moreover, autophagy inhibitor 3‑methyladenine treatment inhibited cell autophagy and apoptosis in miR‑16‑5p‑overexpressing cells. Mechanistic studies demonstrated that miR‑16‑5p could inhibit the BIMP1/NF‑κB signaling pathway and this inhibition was achieved by directly targeting BIMP1. Furthermore, it was found that blockade of the BIMP1/NF‑κB signaling pathway inversed the inhibitory effects of miR‑16‑5p knockdown on autophagy in BC cells. In vivo experiments further verified the tumor‑suppressive effect on BC of the miR‑16‑5p/BIMP1/NF‑κB axis. Therefore, the results of the present study indicated that miR‑16‑5p promotes autophagy of BC cells via the BIMP1/NF‑κB signaling pathway, and an improved understanding of miR‑16‑5p function may provide therapeutic targets for clinical intervention in this disease.

Keywords: NF‑κB signaling pathway; autophagy; bladder cancer; caspase recruitment domain family member 10; microRNA‑16‑5p.

MeSH terms

  • Animals
  • Apoptosis
  • Autophagy*
  • Autophagy-Related Proteins / metabolism*
  • Beclin-1 / metabolism
  • Biomarkers, Tumor
  • CARD Signaling Adaptor Proteins / metabolism
  • Carcinoma / genetics
  • Carcinoma / metabolism
  • Cell Line, Tumor
  • Cell Proliferation
  • Cell Survival
  • Gene Expression Regulation, Neoplastic*
  • Gene Knockdown Techniques
  • Humans
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mice, Nude
  • MicroRNAs / physiology*
  • Microtubule-Associated Proteins / metabolism
  • NF-kappa B / metabolism
  • RNA-Binding Proteins / metabolism
  • Signal Transduction*
  • Urinary Bladder Neoplasms / genetics*
  • Urinary Bladder Neoplasms / metabolism*

Substances

  • Autophagy-Related Proteins
  • Beclin-1
  • Biomarkers, Tumor
  • CARD Signaling Adaptor Proteins
  • CARD10 protein, human
  • MAP1LC3B protein, human
  • MIRN16 microRNA, human
  • MicroRNAs
  • Microtubule-Associated Proteins
  • NF-kappa B
  • P62 protein, human
  • RNA-Binding Proteins

Grants and funding

The present study was supported by grants from the Scientific Research Project of the Education Department of Liaoning Province (grant no. QN2019008), China Medical University's 2018 Youth Support Program (Natural Science; grant no. QGZ2018041), the Shenyang Plan Project of Science and Technology (grant no. F19-112-4-098), the National Key R&D Plan Key Research Projects of Precision Medicine (grant no. 2017YFC0908000) and China Medical University's 2019 Discipline Promotion Program.