RNA Methylation dynamics regulates cancer stemness through modulation of cell migration and cell proliferation in the spheroidal model of TNBC

Altered cellular changes in tumors are regulated by a plethora of mechanisms, including post-transcriptional processes such as the reversible m6A RNA methylation which is shown to be involved in multiple steps of RNA processing thus playing a crucial role in gene expression. The individual roles of the writers, erasers and readers is slowly being unravelled and due to their regulatory role at the post transcriptional level, there are reports of both positive and negative modulatory effects on tumorigenesis. In this study, we have studied the role of one of the m6A writer complex proteins, WTAP and the m6A RNA demethylase FTO (fat mass and obesity associated protein) in 2D and 3D culture systems of triple negative breast cancer cells. Genome edited cell lines where WTAP or FTO levels were downregulated show altered cell proliferation through m6A mediated regulation of cell cycle regulators, and altered cell migration properties through modulation of EMT markers. The 3D spheroidal cultures derived from these genome edited lines revealed a role for m6A regulation in CSC marker expression, thus suggesting a potential oncogenic feedback loop between m6A modulators and the major hallmarks of cancer.