N6-Methyladenosine–Modified KREMEN2 Promotes Tumorigenesis and Malignant Progression of High-Grade Serous Ovarian Cancer

Abstract

High-grade serous ovarian cancer (HGSOC) remains the most lethal female cancer by far. Herein, clinical HGSOC samples had higher N⁶-methyladenosine (m⁶A) modification than normal ovarian tissue, and its dysregulation had been reported to drive aberrant transcription and translation programs. However, Kringle-containing transmembrane protein 2 (KREMEN2) and its m⁶A modification have not been fully elucidated in HGSOC. In this study, the data from the high-throughput messenger RNA (mRNA) sequencing of clinical samples were processed using the weighted correlation network analysis and functional enrichment analysis. Results revealed that KREMEN2 was a driver gene in the tumorigenesis of HGSOC and a potential target of m⁶A demethylase fat-mass and obesity-associated protein (FTO). KREMEN2 and FTO levels were upregulated and downregulated, respectively, and correlation analysis showed a significant negative correlation in HGSOC samples. Importantly, upregulated KREMEN2 was remarkably associated with lymph node metastasis, distant metastasis, peritoneal metastasis, and high International Federation of Gynecology and Obstetrics stage (Ⅲ/Ⅳ), independent of the age of patients. KREMEN2 promoted the growth of HGSOC in vitro and in vivo, which was dependent on FTO. The methylated RNA immunoprecipitation qPCR and RNA immunoprecipitation assays were performed to verify the m⁶A level and sites of KREMEN2. FTO overexpression significantly decreased m⁶A modification in the 3′ and 5′ untranslated regions of KREMEN2 mRNA and downregulated its expression. In addition, we found that FTO-mediated m⁶A modification of KREMEN2 mRNA was recognized and stabilized by the m⁶A reader IGF2BP1 rather than by IGF2BP2 or IGF2BP3. This study highlights the m⁶A modification of KREMEN2 and extends the importance of RNA epigenetics in HGSOC.