Anti-t umor Effects of Recombinant Clostridium α-Toxin on Breast Cancer: An In Vitro and In Vivo Study

Abstract

Cancer is the second leading cause of death worldwide, surpassed only by cardiovascular diseases. This study investigated the anticancer effects of recombinant Clostridium alpha-toxin on breast cancer, both in vitro and in vivo. The entire coding sequence of a codon-optimized alpha- toxin was designed, cloned into the pET28a (+) vector, and expressed as recombinant alpha-toxin in Escherichia coli (E. coli) BL 21(DE3) cells transformed with the recombinant plasmid. The recombinant alpha-toxin was purified using Ni2+ affinity chromatography, and its accuracy and purity were confirmed through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis. The anticancer effects of purified alpha-toxin were then assessed in vitro and animal models against MCF-7 breast cancer cells. Protein analysis confirmed the presence of a 48 kDa band corresponding to the recombinant alpha-toxin. Additionally, the IC50 values of alpha-toxin against MCF-7 cells at 24, 48, and 72 h were 407.3 +/- 2.392 mu g/mL, 287.3 +/- 5.411 mu g/mL, and 258.1 +/- 4.671 mu g/mL, respectively. In vivo, results demonstrated a significant reduction in mean cancer nodule size following alpha-toxin treatment (p<0.001). These findings suggest that alpha-toxin may serve as a promising candidate for breast cancer therapy.