anti-tumor effects of recombinant clostridium α-toxin on breast cancer: an in vitro and in vivo study

Cancer is the second leading cause of death worldwide, surpassed only by cardiovascular diseases. this study investigated the anticancer effects of recombinant clostridium α-toxin on breast cancer, both in vitro and in vivo. the entire coding sequence of a codon-optimized α-toxin was designed, cloned into the pet28a (+) vector, and expressed as recombinant α-toxin in escherichia coli (e. coli) bl 21(de3) cells transformed with the recombinant plasmid. the recombinant α-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 α-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 α-toxin. additionally, the ic₅₀ values of α-toxin against mcf-7 cells at 24, 48, and 72 h were 407.3±2.392 μg/ml, 287.3±5.411 μg/ml, and 258.1±4.671 μg/ml, respectively. in vivo, results demonstrated a significant reduction in mean cancer nodule size following α-toxin treatment (p<0.001). these findings suggest that α-toxin may serve as a promising candidate for breast cancer therapy.