CKS2, regulated by METTL3, contributes to osteosarcoma progression in an IGF2BP1-dependent manner

Emerging evidence implicates N6-methyladenosine (m⁶A) RNA modification in osteosarcoma (OS) pathogenesis, but the downstream effectors and regulatory networks remain not fully elucidated. Cyclin-dependent kinase subunit 2 (CKS2) has been identified as a potential key player in OS. This study investigates its role in OS progression and its regulation via METTL3-mediated m⁶A modification.

Differentially expressed genes (DEGs) were analyzed using the GSE16088 dataset from the GEO database. Candidate targets were validated in OS tissues by qRT-PCR, and the gene with the highest fold change was selected. The role of CKS2 was examined using in vitro and in vivo assays. The relationships among CKS2, METTL3, and IGF2BP1 were analyzed via bioinformatics, correlation analysis, RIP, MeRIP, qRT-PCR, Western blotting, and mRNA stability assays. Rescue experiments explored the functional relationship between METTL3 and CKS2.

CKS2 was identified as one of the most upregulated genes in OS, a finding confirmed in OS tissues. Its silencing suppressed OS cell proliferation, colony formation, migration, invasion, and tumor growth. Furthermore, METTL3 expression was positively associated with CKS2 levels, and METTL3 overexpression increased CKS2 mRNA stability in an m⁶A-dependent manner. Interestingly, IGF2BP1 bound directly to m⁶A-modified CKS2 transcripts and maintained their stability. Functionally, METTL3 overexpression partially rescued the suppressive effects of CKS2 silencing on OS cells.

Our study identified a METTL3/IGF2BP1-CKS2 axis that promotes OS progression via m⁶A-dependent mRNA stabilization, highlighting CKS2 as a potential therapeutic target.