Biochemical mechanism of acetaminophen (APAP) toxicity in human melanocytic melanoma cell lines: a tyrosinase produrg activation approach

摘要: 1289 Purpose: In this work, we investigated the biochemical mechanism of acetaminophen (APAP) induced toxicity in human melanocytic melanoma cell lines. Because tyrosinase is found abundantly it was hypothesized that can be utilized as a molecular target for selective bioactivation APAP, prodrug, to cytotoxic quinone reactive intermediate species, hence resulting cells. Experimental procedures: UV-VIS spectra used investigate enzymatic oxidation APAP by enzyme. GSH depletion measure extent metabolism and rat liver microsomes. The four lines (which express functional tyrosinase), non-melanoma one amelanocytic line do not tyrosinase) using MTT assay. SK-MEL-28 number modulators. Results: Our findings indicate metabolized 87% 33% microsomes measured at 2 h. Ascorbic acid NADH, reducing agents, were significantly depleted during tyrosinase. IC50 (48 h) towards SK-MEL-28, MeWo SK-MEL-5, B16-F0 cells 100 µM which within therapeutic concentration when given regular dose an antipyretic analgesic drug. However, showed no significant BJ, Saos-2, SW-620, PC3 C32 line, thus demonstrating selectively toxic Dicoumarol, diaphorase inhibitor, 1-bromoheptane, depleting agent, enhanced acid, effective preventing toxicity. Trifluoperazine cyclosporin A, inhibitors permeability transition pore mitochondria, prevented caused time dose-dependent decline intracellular concentration, preceded also led ROS formation exacerbated dicoumarol 1-bromoheptane whereas cyslosporin A trifluoperazine it. Conclusion: investigation suggests activated species cells, depletion, mitochondrial contributed APAP’s