Methylated arsenic metabolites bind to PML protein but do not induce cellular differentiation and PML-RARα protein degradation

Abstract

Arsenic trioxide (As₂O₃) is one of the most effective therapeutic agents used for patients with acute promyelocytic leukemia (APL). The probable explanation for As₂O₃-induced cell differentiation is the direct targeting of PML-RARα oncoprotein by As₂O₃, which results in initiation of PML-RARα degradation. However, after injection, As₂O₃ is rapidly methylated in body to different intermediate metabolites such as trivalent monomethylarsonous acid (MMA^III) and dimethylarsinous acid (DMA^III), therefore, it remains unknown that which arsenic specie is actually responsible for the therapeutic effects against APL. Here we have shown the role of As₂O₃ (as iAs^III) and its intermediate metabolites (i.e., MMA^III/DMA^III) in NB4 cells. Inorganic iAs^III predominantly showed induction of cell differentiation, while MMA^III and DMA^III specifically showed to induce mitochondria and endoplasmic reticulum-mediated apoptosis, respectively. On the other hand, in contrast to iAs^III, MMA^III showed stronger binding affinity for ring domain of PML recombinant protein, however, could not induce PML protein SUMOylation and ubiquitin/proteasome degradation. In summary, our results suggest that the binding of arsenicals to the ring domain of PML proteins is not associated with the degradation of PML-RARα fusion protein. Moreover, methylated arsenicals can efficiently lead to cellular apoptosis, however, they are incapable of inducing NB4 cell differentiation.