METTL3-mediated m6A modification of pri-miR-148a-3p affects prostate cancer progression by regulating TXNIP

Environ Toxicol. 2023 Oct;38(10):2377-2390. doi: 10.1002/tox.23874. Epub 2023 Jul 14.

Abstract

Objective: Prostate cancer (PCa) severely affects men's health worldwide. The mechanism of methyltransferase-like 3 (METTL3) in affecting PCa development by regulating miR-148a-3p expression via N6-methyladenosine (m6A) modification was investigated.

Methods: METTL3, miR-148a-3p, and thioredoxin interacting protein (TXNIP) levels were determined using RT-qPCR and Western blotting. The m6A modification level of miR-148a-3p was observed by Me-RIP assay. Bioinformatics website predicted miR-148a-3p and TXNIP levels in PCa and their correlation, and the binding site between them was verified by dual-luciferase assay. The proliferation, migration, invasion, and apoptosis of PCa cells were examined by CCK-8 assay, Transwell assay, and flow cytometry. A transplanted tumor model was established in nude mice to observe the tumor growth ability, followed by determination of TXNIP levels in tumor tissues by immunohistochemistry.

Results: METTL3 interference restrained the proliferation, migration, and invasion and promoted apoptosis of PCa cells. METTL3 up-regulated miR-148a-3p by promoting the m6A modification of pri-miR-148a-3p in PCa cells. miR-148a-3p overexpression nullified the inhibitory actions of silencing METTL3 on PCa cell growth. miR-148a-3p facilitated PCa cell growth by silencing TXNIP. METTL3 interference inhibited tumor growth by down-regulating miR-148a-3p and up-regulating TXNIP.

Conclusion: METTL3 promoted miR-148a-3p by mediating the m6A modification of pri-miR-148a-3p, thereby targeting TXNIP, interfering with METTL3 to inhibit the proliferation, migration and invasion of PCa cells, promote apoptosis, and inhibit tumor growth in nude mice.

Keywords: METTL3; TXNIP; epigenetics; m6A; miR-148a-3p; pri-miR-148a-3p; prostate cancer.

MeSH terms

  • Animals
  • Carrier Proteins / genetics
  • Cell Line, Tumor
  • Cell Proliferation / genetics
  • Humans
  • Male
  • Methyltransferases / genetics
  • Methyltransferases / metabolism
  • Mice
  • Mice, Nude
  • MicroRNAs* / genetics
  • MicroRNAs* / metabolism
  • Prostate
  • Prostatic Neoplasms* / genetics

Substances

  • MicroRNAs
  • METTL3 protein, human
  • Methyltransferases
  • TXNIP protein, human
  • Carrier Proteins