A Bioassay Using the Human Hepatoblastoma Cell Line HepG2 for Detecting Phototoxicity of Furocoumarins

Abstract

We successfully evaluated the human hepatoblastoma cell line HepG2 as a model to assess phototoxicity of coumarins. Five natural furocoumarins were tested and their phototoxic activities, obtained by measuring cell viability in the presence of UV using the MTT test, were as follows: xanthotoxin (8-MOP) >> heraclenol = trichoclin = imperatorin >> peucedanin, both in growing and confluent cell cultures. This easy-to-perform, miniaturised, quantitative and sensitive method could therefore be used as a primary screening test for phototoxicity of a large number of compounds and plant extracts.

HepG2 cell line - phototoxicity - cytotoxicity - furocoumarins

References

• 1 Averbeck D. Recent advances in psoralen phototoxicity mechanism.  Photochemistry and Photobiology. 1989;  50 859-82
• 2 Ashwood-Smith M J, Poulton G A, Ceska O, Liu M, Furniss E. An ultrasensitive bioassay for the detection of furocoumarins and other photosensitizing molecules.  Photochemistry and Photobiology. 1983;  38 113-8
• 3 Van der Sluis W G, Van Arkel J, Fisher F C, Labadie R P. Thin-layer chromatographic assay of photoactive compounds (furocoumarins) using the fungus Penicillium expansum as a test organism.  Journal of Chromatography. 1981;  214 349-59
• 4 Kavli G, Volden G. The Candida test for phototoxicity.  Photodermatology. 1984;  1 204-7
• 5 Tenenbaum S, DiNardo J, Morris W E, Wolf B A, Schnetzinger R W. A quantitative in vitro assay for the evaluation of phototoxic potential of topically applied materials.  Cell Biology and Toxicology. 1984;  1 1-9
• 6 Schimmer O, Beck R, Dietz U. Phototoxicität und Photomutagenität von Furocumarinen und Furocumarindrogen bei Chlamydomonas reinhardii .  Planta Medica. 1980;  40 68-76
• 7 Ojala T, Vuorela P, Kiviranta J, Vuorela H, Hiltunen R. A biossay using Artemia salina for detecting phototoxicity of plant coumarins.  Planta Medica. 1999;  65 715-8
• 8 Abel G, Schimmer O. Mutagenicity and toxicity of furocoumarins: comparative investigations in two test systems.  Mutation Research. 1981;  90 451-61
• 9 Mandula B B, Pathak M A. Metabolic reactions in vitro of psoralens with liver and epidermis.  Biochemical Pharmacology. 1979;  28 127-32
• 10 Dall’Acqua F, Vedaldi D, Baccichetti F, Bordin F, Averbeck D. Photochemotherapy of skin-diseases: comparative studies on the photochemical and photobiological properties of various mono- and bifunctional agents.  Farmaco Edizione Scientifica. 1981;  36 519-35
• 11 Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays.  Journal of Immunological Methods. 1983;  65 55-63
• 12 Carmichael J, DeGraff W G, Gazdar A F, Minna J D, Mitchell J B. Evaluation of tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing.  Cancer Research. 1987;  47 936-42