TNF-α drives bladder cancer metastasis via METTL3-mediated m6A modification to promote CLASP2/IQGAP1-dependent cytoskeleton remodeling

  • Biochim Biophys Acta Mol Basis Dis. 2025 Mar 19;1871(5):167811. doi: 10.1016/j.bbadis.2025.167811.
Lingxiao Chen  1 ,  Cheng He  1 ,  Zhenyu Ou  1 ,  Cheng Zhao  2
Affiliations
  • 1. Department of Urology, Xiangya Hospital, Central South University, Changsha City, Hunan Province, China.
  • 2. Department of Urology, Xiangya Hospital, Central South University, Changsha City, Hunan Province, China. Electronic address: [email protected].
PMID: 40118293 DOI: 10.1016/j.bbadis.2025.167811
Abstract

Background: Bladder Cancer (BCa) metastasis is a multi-step process triggered by Cytoskeleton reorganization. However, the regulation mechanism of Cytoskeleton reorganization in BCa remains ambiguous. This study elucidated the influence of tumor necrosis factor-alpha (TNF-α) in Cytoskeleton remodeling during BCa metastasis and its possible mechanisms.

Methods: Colony formation, scratch, transwell, and the nude mouse model were adopted to evaluate the growth and metastasis. Molecular expression was assessed by immunohistochemical staining, quantitative Real-Time PCR (qRT-PCR), and Western blotting. The N6-methyladenosine (m6A) level was detected by methylated RNA immunoprecipitation (MeRIP). Protein interaction was validated by Co-immunoprecipitation (Co-IP). Immunofluorescence staining was used to identify rearrangement of actin filament fibers (F-actin) and protein colocalization.

Results: TNF-α facilitated cytoplasmic linker associated protein 2 (CLASP2) and methyltransferase like 3 (METTL3) expression in a dose (10-50 ng/mL)-dependent manner in BCa. CLASP2 high expression suggested a shorter overall survival of BCa patients. CLASP2 deficiency suppressed BCa cell proliferation, migration, and invasion via disrupting F-actin Cytoskeleton. Mechanistically, TNF-α promoted METTL3-mediated m6A modification of CLASP2 to enhance CLASP2 mRNA stability. Moreover, CLASP2 directly interplayed with IQ motif containing GTPase activating protein 1 (IQGAP1) to regulate F-actin Cytoskeleton remodeling. In vivo data demonstrated that inhibition of METTL3/CLASP2 axis delayed lung metastasis in nude mice.

Conclusion: TNF-α favors BCa cell metastasis, which involves METTL3-mediated m6A modification of CLASP2 that interacts with IQGAP1, thus leading to F-actin Cytoskeleton remodeling. Our findings suggest targeting TNF-α/METTL3/CLASP2/IQGAP1 axis as a potential avenue for promising treatment for BCa.

Keywords
Bladder cancer; CLASP2; Cytoskeleton reorganization; IQGAP1; METTL3; TNF-α.
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