Sulforaphene suppressed cell proliferation and promoted apoptosis of COV362 cells in endometrioid ovarian cancer

PeerJ. 2023 Nov 21:11:e16308. doi: 10.7717/peerj.16308. eCollection 2023.

Abstract

Aim: N6-methyladenosine (m6A) RNA methylation exerts a regulatory effect on endometrioid ovarian cancer (EOC), but the specific m6A regulator genes in EOC remain to be explored. This study investigated that sulforaphene (Sul) is implicated in EOC development by regulating methyltransferase-like 3 (METTL3).

Methods: The dysregulated m6A RNA methylation genes in EOC were determined by methylated RNA immunoprecipitation (MeRIP-seq) and RNA sequencing. The roles of METTL3 and/or Sul on viability, proliferative ability, cell cycle, and apoptosis of EOC cells were determined by MTT, colony formation, flow cytometry, and TUNEL staining assay, respectively. The expression of METTL3 and apoptosis-related proteins in EOC cells was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot assays.

Results: Five m6A RNA methylation regulators (METTL3, ELF3, IGF2BP2, FTO, and METTL14) were differentially expressed in EOC, among which METTL3 had the highest expression level. Silencing METTL3 reduced the clonal expansion and viability of EOC cells, and caused the cells to arrest in the G0/G1 phase. This also promoted apoptosis in the EOC cells and activated the FAS/FADD and mitochondrial apoptosis pathways. In contrast, overexpressing METTL3 had the opposite effect. Sul, in a dose-dependent manner, reduced the viability of EOC cells but promoted their apoptosis. Sul also increased the levels of IGF2BP2 and FAS, while decreasing the levels of KRT8 and METTL3. Furthermore, Sul was able to reverse the effects of METTL3 overexpression on EOC cells.

Conclusions: Sul could suppress cell proliferation and promote apoptosis of EOC cells by inhibiting the METTL3 to activate the FAS/FADD and apoptosis-associated pathways.

Keywords: Endometrioid ovarian cancer; FAS; FASL; Methyltransferase-like 3; Sulforaphane.

MeSH terms

  • Alpha-Ketoglutarate-Dependent Dioxygenase FTO
  • Apoptosis / genetics
  • Carcinoma, Endometrioid* / genetics
  • Carcinoma, Ovarian Epithelial
  • Cell Proliferation / genetics
  • Female
  • Humans
  • Methyltransferases / genetics
  • Ovarian Neoplasms* / genetics
  • RNA
  • RNA-Binding Proteins

Substances

  • sulphoraphene
  • RNA
  • METTL3 protein, human
  • Methyltransferases
  • IGF2BP2 protein, human
  • RNA-Binding Proteins
  • FTO protein, human
  • Alpha-Ketoglutarate-Dependent Dioxygenase FTO

Grants and funding

This research was funded by the Public Welfare Technology Project of Zhejiang Province (LGF19H160011) and the Zhejiang Provincial Health and Medicine Science and Technology Project (2019ZD001). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.