This study aimed to examine how mesoporous silica nanoparticles-chitosan-folic acid impacted the release of recombinant Azurin within the tumor environment. The goal was to trigger apoptosis and stimulate immune responses against both transformed and normal cells in BALB/c mice. The study found that the use of rAzu-MSNs-CS-FA, a specific formulation containing mesoporous silica nanoparticles-chitosan-folic acid, resulted in pH-responsive behavior and slower release of rAzurin compared to other groups. This formulation inhibited MCF7 cells at higher concentrations, induced apoptosis in cells, and caused DNA degradation. It also increased the uptake efficiency of rAzurin and stimulated the secretion of TNF-α, INF-γ, and IL-4 while inhibiting the secretion of IL-6. Furthermore, it regulated the expression of specific genes (upregulating tlr3 and downregulating tlr2, 4, and 9). In animal studies with BALB/c mice, the rAzu-MSNs-CS-FA formulation led to tumor regression and decreased tumor volume over 21 days. Overall, this formulation showed promising results in inducing cytotoxic effects against cancer cells, promoting apoptosis, and eliciting appropriate immune responses, suggesting its potential as a valuable therapy for breast cancer.