SB431542, a Selective Inhibitor of the TGF-β Type I Receptor, Enhances Doxorubicin Antitumor Activity via p63 Activation in Mutant p53 Breast Cancer Cells.

BACKGROUND

TP53 gene mutations are common in breast cancer and are linked to chemoresistance. p63, a p53 family member, can induce apoptosis independently of p53, representing a potential therapeutic target in TP53-mutant tumors. This study evaluated the synergistic effects of SB431542, a TGF-β type I receptor inhibitor, and doxorubicin in TP53-mutant breast cancer cells.

METHODS

Isoform-specific RT-PCR was used to assess TAp63 and ΔNp63 expression following SB431542 treatment in T47D, MDA-MB-231, and MDA-MB-468 cells. Cell viability was assessed using the CCK8 assay. Synergistic interaction was quantified using the Coefficient of Drug Interaction (CDI). Caspase-3/7 activity assays and immunocytochemistry analyses were performed to evaluate apoptotic signaling and p63 expression. Inhibition studies using PETα, a p53-family inhibitor, and a pan-caspase inhibitor were conducted to determine the pathway dependency of the observed effects.

RESULTS

SB431542 selectively increased TAp63 but not ΔNp63 expression in all three TP53-mutant breast cancer cells. GAS6, a TAp63 target, was also upregulated by SB431542. Treatment with SB431542 and doxorubicin used in combination significantly reduced cell viability (CDI 0.54-0.63), increased caspase activity, and enhanced p63 expression. The anticancer effect was significantly reduced by co-treatment with either the p53-family inhibitor or the pan-caspase inhibitor, confirming that the cytotoxic response was mediated through TAp63 and caspase activation.

CONCLUSIONS

SB431542 potentiates doxorubicin-induced apoptosis in TP53-mutant breast cancer cells by upregulating TAp63 and activating caspase-dependent pathways. These findings suggest that targeting the TGF-β/TAp63 signaling axis may offer a novel therapeutic approach to overcome chemoresistance in aggressive, TP53-mutant breast cancers.