Document Type : Original Article
Authors
1
Department of Microbiology and Immunology, School of Medicine, Kashan University of Medical Sciences, Kashan, Iran.
2
Department of Bacteriology and Virology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan
3
Dental and Periodontal Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
4
Infectious and Tropical Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
5
Skin Diseases and Leishmaniasis Research Center, Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Abstract
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.
Keywords