Vitamin C and K3 combination causes enhanced anticancer activity against RT-4 bladder cancer cells

Introduction: Vitamin C (VC), Vitamin K₃ (VK₃) and the combination (VC:VK₃) were evaluated against human bladder cancer cell lines RT-4 and T24 to evaluate their synergistic anticancer activity. Methods/Results: An MTT assay compared a 1hr pulsed versus a 5hr continuous exposure. VC:VK₃ was synergistic, increasing the antitumor activity 12-to 24 fold for RT-4 cells. VC:VK₃ pulsed versus continuous exposure produced comparable CD₅₀ values, indicating a triggered response involving a catalase reversible redox mechanism generating hydrogen peroxide. Hydrogen peroxide production caused lipid peroxidation and depletion of cellular thiols. ATP levels were measured over 5hrs to determine metabolic effects where VC:VK₃ caused a unique spike in ATP levels. Though the cause of the ATP spike is unknown a possible mechanism is a shunt formed around a defective region of complex III of the ETC from coenzyme Q to cytochrome c, producing a shift from glycolytic to oxidative metabolism and a diminution of lactic acidosis. Analysis of mitochondrial and extra mitochondrial calcium levels revealed a unique calcium pattern for RT4 cells treated with CD₉₀ doses of VC, VK₃ or VC:VK₃. Conclusion: VC:VK₃ was able to cause autoschizic cell death through oxidative stress, thiol depletion, lipid peroxidation, modification of ATP levels and calcium regulation. Because of these results, VC:VK₃ was granted orphan drug status for the treatment of metastatic or locally advanced, inoperable transitional cell carcinoma of the urothelium (stage III and IV bladder cancer). Efforts are underway to conduct a phase II clinical trial for this indication.