Inotilone suppresses fibronectin 1 expression and inhibits the growth of triple-negative breast cancer xenografts under prolonged nicotine exposure

Breast cancer is one of the most significant cancers affecting women. The link between smoking and breast cancer development, along with its effects on treatment, needs further investigation for clear validation. Our previous study using breast cancer patient-derived xenograft (PDX) models revealed that prolonged nicotine exposure accelerates HER2-positive tumor progression and reduces trastuzumab efficacy. In this study, RNA sequencing of nicotine-exposed PDX tumors revealed the 50 most altered differentially expressed genes (DEGs), with PPI networks analyzed via STRING. Higher fibronectin (FN1) expression in cancer cells is linked to worse survival in triple-negative breast cancer (TNBC) patients (**p < 0.01). To investigate the effects of nicotine on aggressive breast cancer, our study utilized the TNBC-PDX and MDA-MB-231 cell models. Our results revealed that prolonged nicotine exposure promoted tumor growth, increased FN1 protein levels, enhanced cell adhesion and metastasis-related proteins, and diminished the efficacy of Cisplatin. Our previous research emphasized the therapeutic potential of natural compounds in combating nicotine-induced breast cancer. Prolonged nicotine (10 mM) exposure for 17 generations (p17), followed by 24-hour treatment with 15 μM Inotilone, reduced FN1 protein expression and significantly inhibited MDA-MB-231 cell migration (**p < 0.01). To validate the tumor-suppressive effects of Inotilone and its inhibition of nicotine-induced FN1 overexpression beyond TNBC cells, we utilized a HER2 + HCC-1954 breast cancer xenograft model in NSG mice, which showed significantly reduced tumor growth upon intraperitoneal Inotilone treatment (40 mg/kg every four days) compared to controls (**p < 0.01). In conclusion, Inotilone demonstrates promise as an adjunctive therapy for breast cancer by effectively downregulating FN1 expression and mitigating tumor progression associated with prolonged nicotine exposure in vivo.