Rezaei Khozani Nahid, Shayesteh Pour Mohammad, Yekani Mina, Hejazi Seyed Hossein, Saffari Mahmood
Department of Microbiology and Immunology, School of Medicine, Kashan University of Medical Sciences, Kashan, Iran.
Department of Bacteriology and Virology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Int J Mol Cell Med. 2024;13(4):404-416. doi: 10.22088/IJMCM.BUMS.13.4.404.
Cancer is the second leading cause of death worldwide, surpassed only by cardiovascular diseases. This study investigated the anticancer effects of recombinant α-toxin on breast cancer, both . The entire coding sequence of a codon-optimized was designed, cloned into the pET28a (+) vector, and expressed as recombinant α-toxin in () 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 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. results demonstrated a significant reduction in mean cancer nodule size following α-toxin treatment (<0.001). These findings suggest that α-toxin may serve as a promising candidate for breast cancer therapy.
癌症是全球第二大死因，仅次于心血管疾病。本研究调查了重组α-毒素对乳腺癌的抗癌作用。设计了密码子优化后的完整编码序列，将其克隆到pET28a(+)载体中，并在转化了重组质粒的大肠杆菌(Escherichia coli) BL21(DE3)细胞中表达为重组α-毒素。使用镍离子亲和层析法纯化重组α-毒素，并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质免疫印迹分析确认其准确性和纯度。然后在体外和动物模型中评估纯化的α-毒素对MCF-7乳腺癌细胞的抗癌作用。蛋白质分析证实存在一条与重组α-毒素相对应的48 kDa条带。此外，α-毒素在24、48和72小时对MCF-7细胞的IC₅₀值分别为407.3±2.392 μg/mL、287.3±5.411 μg/mL和258.1±4.671 μg/mL。结果表明，α-毒素治疗后平均癌结节大小显著减小(<0.001)。这些发现表明，α-毒素可能是一种有前景的乳腺癌治疗候选药物。
