title: Correlation between luminescence intensity and cytotoxicity in cell-based cytotoxicity assay using luciferase.
abstract: The luciferase reporter assay has become one of the conventional methods for cytotoxicity evaluation. Typically, the decrease of luminescence expressed by a constitutive promoter is used as an index of cytotoxicity. However, to our knowledge, there have been no reports of the correlation between cytotoxicity and luminescence intensity. In this study, to accurately verify the correlation between them, beetle luciferase was stably expressed in human hepatoma HepG2 cells harboring the multi-integrase mouse artificial chromosome vector. We showed that the cytotoxicity assay using luciferase does not depend on the stability of luciferase protein and the kind of constitutive promoter. Next, HepG2 cells in which green-emitting beetle luciferase was expressed under the control of CAG promoter were exposed to 58 compounds. The luminescence intensity and cytotoxicity curves of cells exposed to 48 compounds showed similar tendencies, whereas those of cells exposed to 10 compounds did not do so, although the curves gradually approached each other with increasing exposure time. Finally, we demonstrated that luciferase expressed under the control of a constitutive promoter can be utilized both as an internal control reporter for normalizing a test reporter and for monitoring cytotoxicity when two kinds of luciferases are simultaneously used in the cytotoxicity assay.
authors.affiliation: Hatano Research Institute, Food and Drug Safety Center, Hadano, Kanagawa 257-8523, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Wakuri
authors.affiliation: Hatano Research Institute, Food and Drug Safety Center, Hadano, Kanagawa 257-8523, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Yamakage
authors.affiliation: Chromosome Engineering Research Center, Tottori University, Yonago, Tottori 683-8503, Japan; Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University, Tottori 683-8503, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Kazuki
authors.affiliation: Chromosome Engineering Research Center, Tottori University, Yonago, Tottori 683-8503, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Kazuki
authors.affiliation: Chromosome Engineering Research Center, Tottori University, Yonago, Tottori 683-8503, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Oshimura
authors.affiliation: Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Aomi, Tokyo 135-0064, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Aburatani
authors.affiliation: Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, Kagawa 761-0395, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Yasunaga
authors.affiliation: Chromosome Engineering Research Center, Tottori University, Yonago, Tottori 683-8503, Japan; Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, Kagawa 761-0395, Japan. Electronic address: y-nakajima@aist.go.jp.
authors.forename: Y
authors.initials: Y
authors.lastname: Nakajima
mesh_terms: D000818:Animals; D022221:Chromosomes, Artificial, Mammalian / Q000235:genetics / Q000378:metabolism; D001517:Coleoptera; D056945:Hep G2 Cells; D006801:Humans; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D051379:Mice; D011401:Promoter Regions, Genetic; D018675:Toxicity Tests / Q000379:methods
chemical_list: D019476:Insect Proteins; D008156:Luciferases
keywords: Cytotoxicity; Internal control reporter; Luciferase; Luciferase reporter assay