Depletion of the m6A demethylases FTO and ALKBH5 impairs growth and metastatic capacity through EMT phenotype change in clear cell renal cell carcinoma

Wei Hu; Niklas Klümper; Doris Schmidt; Manuel Ritter; Jörg Ellinger; Stefan Hauser

Depletion of the m⁶A demethylases FTO and ALKBH5 impairs growth and metastatic capacity through EMT phenotype change in clear cell renal cell carcinoma

Am J Transl Res. 2023 Mar 15;15(3):1744-1755. eCollection 2023.

Authors

Wei Hu^{1

2}, Niklas Klümper¹, Doris Schmidt¹, Manuel Ritter¹, Jörg Ellinger¹, Stefan Hauser¹

Affiliations

¹ Department of Urology, University Hospital Bonn Bonn, Germany.
² Department of Urology, Renmin Hospital, Wuhan University Wuhan, Hubei, China.

PMID: 37056835
PMCID: PMC10086911

Abstract

Background: N⁶-methyladenosine (m⁶A) is one of the most common RNA modifications in eukaryotes and has effects on RNA structure and stability. Recent studies have shown that m⁶A methylation is involved in human carcinogenesis. In the present study, we investigated the effects of m⁶A demethylases FTO and ALKBH5 on renal cell carcinoma (RCC) cell lines.

Methods: The epithelial-mesenchymal in vitro knockdowns of FTO and ALKBH5 induced by antisense oligonucleotides (LNA-GapmeR system) were established in RCC cell lines. Their effects on migration and proliferation were investigated subsequently. The influence of FTO and ALKBH5 knockdown on key epithelial-mesenchymal transition (EMT) genes was analyzed.

Results: Inactivation of FTO and ALKBH5 resulted in decreased proliferation and motility in all cell lines examined (ACHN, Caki-1, 769-P). Vimentin (VIM) was downregulated after the knockdown of FTO and ALKBH5, indicating an EMT switch.

Conclusions: Knockdown of the m⁶A erasers FTO and ALKBH5 inhibits the malignant potential in the cell cultures studied by means of an EMT switch.

Keywords: ALKBH5; FTO; clear cell renal cell carcinoma (ccRCC); epithelial-mesenchymal transition (EMT); m6A-erasers.