The Effect of Riboflavin and Blue Laser Irradiation on B16-F10 Melanoma Cell Death
Aigul K. Gilmutdinova
Federal State Budgetary Educational Institution of Higher Education “Ulyanovsk State University”, Russian Federation
Elena V. Iurova
(Login required)
Federal State Budgetary Educational Institution of Higher Education “Ulyanovsk State University”, Russian Federation
Paper #9451 received 11 Dec 2025; revised manuscript received 03 Apr 2026; accepted for publication 01 May 2026; published online 30 Jun 2026
DOI: 10.18287/JBPE26.12.020312
Abstract
Riboflavin, a natural photosensitizer, is of significant interest for the development of low-toxicity photodynamic therapy (PDT) protocols. This study is focused on the evaluation of the cytotoxic effect and ionic imbalances in B16-F10 melanoma cells induced by riboflavin when irradiating with blue laser light (450 nm wavelength). The dynamics of cell death and changes in intracellular sodium, potassium, and calcium ion concentrations were analyzed with the use of fluorescence microscopy. At a riboflavin concentration of 50 µM under laser irradiation, the signal corresponding to early apoptotic features in B16-F10 cells increased by 2.2 times compared to the control group. The maximum increase associated with late apoptosis and necrosis was observed 6 h after exposure and exceeded the values of the control group by 1.61 times, whereas under combined treatment this effect was detected as early as 1 h after exposure (1.32 times relative to the control group). Analysis of ion homeostasis revealed that riboflavin treatment, particularly in combination with laser irradiation, led to an increase in intracellular Ca2⁺ levels (up to 1.9-fold relative to the control at 6 h after exposure). This was accompanied by an increase in Na⁺ levels and a decrease in K⁺ levels, with minimal K⁺ values observed at 6 h after exposure (approximately 0.6 of the control level). Overall, the results demonstrate the photosensitizing activity of riboflavin and support its further investigation in photodynamic approaches for melanoma. The observed effects provide a basis for further investigation of riboflavin-mediated PDT in combination with other anticancer agents to enhance therapeutic efficacy.
Keywords
Full Text:
PDFReferences
1. R. Kaur, A. Bhardwaj, and S. Gupta, “Cancer treatment therapies: traditional to modern approaches to combat cancers,” Molecular Biology Reports 50(11), 9663–9676 (2023).
2. D. T. Michaeli, J. C. Michaeli, and T. Michaeli, “Advances in cancer therapy: clinical benefit of new cancer drugs,” Aging 15(12), 5232–5234 (2023).
3. Y. Allamyradov, J. Ben Yosef, B. Annamuradov, M. Ateyeh, C. Street, H. Whipple, and A. O. Er, “Photodynamic therapy review: past, present, future, opportunities and challenges,” Photochem 4(4), 434–461 (2024).
4. M. Insińska-Rak, M. Sikorski, and A. Wolnicka-Glubisz, “Riboflavin and its derivates as potential photosensitizers in the photodynamic treatment of skin cancers,” Cells 12(18), 2304 (2023).
5. P. Carpentier, F. Pierrel, N. Duraffourg, B. Guigliarelli, M. Hajj Chehade, L. Flandrin, C. Basset, C. Caux, S. Torelli, and V. Nivière, “Studies of the membrane-bound flavocytochrome MsrQ flavin mononucleotide ( FMN )-binding site reveal an unexpected ubiquinone cofactor,” The FEBS Journal 292(20), 5491–5505 (2025).
6. A. Okamoto, S. Kalathil, X. Deng, K. Hashimoto, R. Nakamura, and K. H. Nealson, “Cell-secreted flavins bound to membrane cytochromes dictate electron transfer reactions to surfaces with diverse charge and pH,” Scientific Reports 4(1), 5628 (2014).
7. M. Insińska-Rak, M. Sikorski, “Riboflavin interactions with oxygen-a survey from the photochemical perspective,” Chemistry – A European Journal 20(47), 15280–15291 (2014).
8. M. Hsieh, S.-K. Hsu, T.-Y. Liu, C.-Y. Wu, and C.-C. Chiu, “Melanoma biology and treatment: a review of novel regulated cell death-based approaches,” Cancer Cell International 24(1), 63 (2024).
9. G. Gunaydin, M. E. Gedik, and S. Ayan, “Photodynamic therapy—current limitations and novel approaches,” Frontiers in Chemistry 9, 691697 (2021).
10. M.-Y. Yang, C.-J. Chang, and L.-Y. Chen, “Blue light induced reactive oxygen species from flavin mononucleotide and flavin adenine dinucleotide on lethality of HeLa cells,” Journal of Photochemistry and Photobiology B: Biology 173, 325–332 (2017).
11. A. V. Juarez, L. D. V. Sosa, A. L. De Paul, A. P. Costa, M. Farina, R. B. Leal, A. I. Torres, and P. Pons, “Riboflavin acetate induces apoptosis in squamous carcinoma cells after photodynamic therapy,” Journal of Photochemistry and Photobiology B: Biology 153, 445–454 (2015).
12. A. Wolnicka-Glubisz, A. Pawlak, M. Insinska-Rak, and A. Zadlo, “Analysis of photoreactivity and phototoxicity of riboflavin’s analogue 3MeTARF,” Journal of Photochemistry and Photobiology B: Biology 205, 111820 (2020).
13. R. A. Akasov, N. V. Sholina, D. A. Khochenkov, A. V. Alova, P. V. Gorelkin, A. S. Erofeev, A. N. Generalova, and E. V. Khaydukov, “Photodynamic therapy of melanoma by blue-light photoactivation of flavin mononucleotide,” Scientific Reports 9(1), 9679 (2019).
14. S. Kacar, C. Hacioglu, and F. Kar, “Irradiated riboflavin over nonradiated one: Potent antimigratory, antiproliferative and cytotoxic effects on glioblastoma cells,” Journal of Cellular and Molecular Medicine 28(8), e18288 (2024).
15. D. L. Sai, J. Lee, D. L. Nguyen, and Y.-P. Kim, “Tailoring photosensitive ROS for advanced photodynamic therapy,” Experimental & Molecular Medicine 53(4), 495–504 (2021).
16. D. Machado, S. M. Shishido, K. C. S. Queiroz, D. N. Oliveira, A. L. C. Faria, R. R. Catharino, C. A. Spek, and C. V. Ferreira, “Irradiated riboflavin diminishes the aggressiveness of melanoma in vitro and in vivo,” PLoS ONE 8(1), e54269 (2013).
17. P. Agostinis, K. Berg, K. A. Cengel, T. H. Foster, A. W. Girotti, S. O. Gollnick, S. M. Hahn, M. R. Hamblin, A. Juzeniene, D. Kessel, M. Korbelik, J. Moan, P. Mroz, D. Nowis, J. Piette, B. C. Wilson, and J. Golab, “Photodynamic therapy of cancer: An update,” CA: A Cancer Journal for Clinicians 61(4), 250–281 (2011).
18. C. Saenz, M. Ethirajan, E. C. Tracy, M.-J. Bowman, J. Cacaccio, T. Ohulchanskyy, H. Baumann, and R. K. Pandey, “Charged groups on pyropheophorbide-based photosensitizers dictate uptake by tumor cells and photodynamic therapy efficacy,” Journal of Photochemistry and Photobiology B: Biology 227, 112375 (2022).
19. N. Clemente, S. Baroni, S. Fiorilla, F. Tasso, S. Reano, C. Borsotti, M. R. Ruggiero, E. Alchera, M. Corrazzari, G. Walker, A. Follenzi, S. G. Crich, and R. Carini, “Boosting intracellular sodium selectively kills hepatocarcinoma cells and induces hepatocellular carcinoma tumor shrinkage in mice,” Communications Biology 6, 574 (2023).
20. F. Lang, E. K. Hoffmann, “Role of ion transport in control of apoptotic cell death,” Comprehensive Physiology 2(3), 2037–2061 (2012).
21. C. D. Bortner, J. A. Cidlowski, “Cell shrinkage and monovalent cation fluxes: Role in apoptosis,” Archives of Biochemistry and Biophysics 462(2), 176–188 (2007).
22. H. Zhang, Z. Chen, A. Zhang, A. A. Gupte, and D. J. Hamilton, “The role of calcium signaling in melanoma,” International Journal of Molecular Sciences 23(3), 1010 (2022).
23. W.-H. Chan, “Photodynamic treatment induces an apoptotic pathway involving calcium, nitric oxide, p53, p21-activated kinase 2, and c-Jun N-terminal kinase and inactivates survival signal in human umbilical vein endothelial cells,” International Journal of Molecular Sciences 12(2), 1041–1059 (2011).
24. N. Sobhani, A. A. Samadani, “Implications of photodynamic cancer therapy: an overview of PDT mechanisms basically and practically,” Journal of the Egyptian National Cancer Institute 33(1), 34 (2021).
Сontact
34 Moskovskoe shosse, Samara, 443086, Russian Federation
Email: j-bpe@ssau.ru
Phone: +7-846-267-4550
© 2014-2025 J-BPE