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CancerGenetics

• Evaluation of the Therapeutic Potential of a Bifidobacterium longum-Derived Postbiotic in Skin Cancer: Concurrent Assessment of Anticancer and Antioxidant Activities

• mohamad javad farhangi,^1,* Behnam Rafiee,² Faezeh Sabetbirjandi,³ Mobina Gholam Fakhrabadi,⁴ Bahman Khameneh Bagheri ,⁵
  1. Department of Biology, Faculty of Basic Sciences, Islamic Azad University of Mashhad, Mashhad, Iran
  2. Department of Animal Science, College of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
  3. Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
  4. Department of Biology, Faculty of Basic Sciences, Islamic Azad University of Mashhad, Mashhad, Iran
  5. Department of Pharmaceutical Control, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
  
  
  
• Introduction: Melanoma is one of the most aggressive forms of skin cancer, characterized by rapid progression and a high potential for metastasis. Oxidative stress and disruption of cellular homeostasis play important roles in melanoma development and progression, highlighting the need for novel therapeutic strategies with improved biological activity and safety. In recent years, increasing attention has been directed toward the therapeutic potential of beneficial microorganisms and their bioactive metabolites. Postbiotics, which comprise non-viable microbial cells, cellular components, and biologically active metabolites, have emerged as promising agents with antioxidant, anti-inflammatory, antimicrobial, and anticancer properties. Among beneficial bacteria, Bifidobacterium longum is recognized for its ability to produce diverse bioactive metabolites that may modulate cellular functions and disease-related pathways. However, the potential anticancer and antioxidant effects of B. longum-derived postbiotics against melanoma remain insufficiently characterized. Therefore, this study aimed to evaluate the therapeutic potential of B. longum-derived postbiotics by investigating their cytotoxic effects against B16F10 murine melanoma cells and their antioxidant activity using the DPPH radical-scavenging assay.
• Methods: Bifidobacterium longum was cultured in de Man–Rogosa–Sharpe (MRS) broth supplemented with a prebiotic substrate. Following the fermentation process, the culture was processed to obtain a cell-free postbiotic preparation derived from B. longum. The obtained postbiotic was subsequently evaluated for its anticancer and antioxidant activities. The anticancer activity of the postbiotic was investigated against the murine melanoma cell line B16F10 using the MTT assay. B16F10 cells were exposed to different concentrations of the postbiotic for the specified incubation period. Cell viability was determined based on the reduction of MTT to formazan crystals, and absorbance was measured at 570 nm. Cell viability was calculated as a percentage relative to the untreated control. The antioxidant activity of the postbiotic was assessed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging assay. Different concentrations of the postbiotic were incubated with DPPH solution, and the absorbance was measured at 517 nm. Vitamin C (ascorbic acid) was used as a positive control. The antioxidant activity was expressed as the percentage of DPPH radical-scavenging activity compared with the control. All experiments were performed in independent replicates, and the data were subjected to appropriate statistical analysis.
• Results: The MTT assay demonstrated that the postbiotic derived from Bifidobacterium longum exerted a considerable cytotoxic effect on B16-F10 cells. Treatment with the B. longum postbiotic at concentrations of 0.125, 0.25, and 0.5 reduced cell viability to 68.70 ± 4.36%, 34.52 ± 3.54%, and 29.56 ± 2.96%, respectively, compared with the untreated control. The corresponding cytotoxicity rates were 31.30%, 65.48%, and 70.44%, respectively. The highest cytotoxic effect was observed at a concentration of 0.5. Based on the concentration–response data, the IC₅₀ of the B. longum postbiotic was estimated to be approximately 0.18. These findings suggest that the B. longum postbiotic possesses promising cytotoxic potential against B16-F10 cells. The DPPH radical scavenging assay demonstrated that the tested Langum sample exhibited considerable free-radical scavenging activity. The radical inhibition rates at concentrations of 0.0625, 0.25, and 0.5 were 21.11%, 33.87%, and 64.48%, respectively. In comparison, vitamin C, used as the positive control, showed 53.14% radical scavenging activity. The antioxidant activity of the sample increased with increasing concentration, with the highest scavenging activity observed at a concentration of 0.5. These findings indicate a considerable free-radical scavenging capacity of the Langum sample and suggest its potential antioxidant activity.
• Conclusion: Overall, the findings of this study demonstrated that the postbiotic derived from Bifidobacterium longum exhibits considerable antioxidant activity and concentration-dependent cytotoxic effects against B16-F10 cells. The increased free-radical scavenging activity observed in the DPPH assay, together with the marked reduction in B16-F10 cell viability in the MTT assay, highlights the promising biological potential of the B. longum postbiotic. These findings suggest that B. longum postbiotic may represent a promising candidate for further investigation as a bioactive compound with potential complementary anticancer applications. However, further in vitro and in vivo studies are required to validate these findings and elucidate the underlying molecular mechanisms.
• Keywords: Bifidobacterium longum-Mtt-Dpph-antioxidant-anticancer