High-throughput genotoxicity assay identifies antioxidants as inducers of DNA damage response and cell death

Human ATAD5 is a biomarker for identifying genotoxic compounds because ATAD5 protein levels increase posttranscriptionally in response to DNA damage. We screened over 4,000 compounds with a cell-based quantitative high-throughput ATAD5-luciferase assay detecting genotoxic compounds. We identified 22 antioxidants, including resveratrol, genistein, and baicalein, that are currently used or investigated for the treatment of cardiovascular disease, type 2 diabetes, osteopenia, osteoporosis, and chronic hepatitis, as well as for antiaging. Treatment of dividing cells with these compounds induced DNA damage and resulted in cell death. Despite their genotoxic effects, resveratrol, genistein, and baicalein did not cause mutagenesis, which is a major side effect of conventional anticancer drugs. Furthermore, resveratrol and genistein killed multidrug-resistant cancer cells. We therefore propose that resveratrol, genistein, and baicalein are attractive candidates for improved chemotherapeutic agents.

[1]  A. Dutra,et al.  DNA damage responses by human ELG1 in S phase are important to maintain genomic integrity , 2009, Cell cycle.

[2]  C. Dabrosin,et al.  Resveratrol induces apoptosis and inhibits angiogenesis in human breast cancer xenografts in vivo. , 2006, Cancer letters.

[3]  Nancy F. Hansen,et al.  Predisposition to Cancer Caused by Genetic and Functional Defects of Mammalian Atad5 , 2011, PLoS genetics.

[4]  A. Bitto,et al.  Effects of the phytoestrogen genistein on some predictors of cardiovascular risk in osteopenic, postmenopausal women: a two-year randomized, double-blind, placebo-controlled study. , 2007, The Journal of clinical endocrinology and metabolism.

[5]  K. Tanikawa,et al.  A multicenter randomized controlled clinical trial of Shosaiko-to in chronic active hepatitis , 1989, Gastroenterologia Japonica.

[6]  T. Frank,et al.  Study on the Pharmacokinetics of Synthetic Genistein after Multiple Oral Intake in Post-menopausal Women , 2009, Arzneimittelforschung.

[7]  Ruili Huang,et al.  Compound Cytotoxicity Profiling Using Quantitative High-Throughput Screening , 2007, Environmental health perspectives.

[8]  Jerzy Jankun,et al.  In vivo and in vitro effect of baicalein on human prostate cancer cells. , 2005, International journal of oncology.

[9]  M. Mann,et al.  Regulation of translesion synthesis DNA polymerase eta by monoubiquitination. , 2010, Molecular cell.

[10]  J. Barrett,et al.  Cell‐transforming activity and mutagenicity of 5 phytoestrogens in cultured mammalian cells , 2003, International journal of cancer.

[11]  M. Nihal,et al.  Resveratrol-caused apoptosis of human prostate carcinoma LNCaP cells is mediated via modulation of phosphatidylinositol 3′-kinase/Akt pathway and Bcl-2 family proteins , 2006, Molecular Cancer Therapeutics.

[12]  N. Miyata,et al.  Resveratrol, a naturally occurring polyphenol, induces sister chromatid exchanges in a Chinese hamster lung (CHL) cell line. , 2001, Mutation research.

[13]  F. Lu,et al.  Baicalein, a Novel Apoptotic Agent for Hepatoma Cell Lines:A Potential Medicine for Hepatoma , 2000, Nutrition and cancer.

[14]  Christopher P Austin,et al.  Characterization of chemical libraries for luciferase inhibitory activity. , 2008, Journal of medicinal chemistry.

[15]  H. Stopper,et al.  Genotoxicity of phytoestrogens. , 2005, Mutation research.

[16]  K. Myung,et al.  Suppression of gross chromosomal rearrangements by a new alternative replication factor C complex. , 2007, Biochemical and biophysical research communications.

[17]  R. Mehta,et al.  Genistein induces maturation of cultured human breast cancer cells and prevents tumor growth in nude mice. , 1998, The American journal of clinical nutrition.

[18]  Robert E. Johnson,et al.  hRAD30 mutations in the variant form of xeroderma pigmentosum. , 1999, Science.

[19]  K. Myung,et al.  Dynamic regulation of PCNA ubiquitylation/deubiquitylation , 2011, FEBS letters.

[20]  I. Pastan,et al.  Isolation and genetic characterization of human KB cell lines resistant to multiple drugs , 1985, Somatic cell and molecular genetics.

[21]  S. Bao,et al.  Genistein induces G2/M cell cycle arrest and apoptosis of human ovarian cancer cells via activation of DNA damage checkpoint pathways , 2009, Cell biology international.

[22]  E. Friedberg,et al.  Trading places: how do DNA polymerases switch during translesion DNA synthesis? , 2005, Molecular cell.

[23]  Juan-juan Chen,et al.  Anticancer activity of resveratrol on implanted human primary gastric carcinoma cells in nude mice. , 2005, World journal of gastroenterology.

[24]  Robert E. Johnson,et al.  Efficient bypass of a thymine-thymine dimer by yeast DNA polymerase, Poleta. , 1999, Science.

[25]  A. Bitto,et al.  Effects of the Phytoestrogen Genistein on Bone Metabolism in Osteopenic Postmenopausal Women , 2007, Annals of Internal Medicine.

[26]  L. Ferguson.  Role of plant polyphenols in genomic stability. , 2001, Mutation research.

[27]  Ruili Huang,et al.  Characterization of environmental chemicals with potential for DNA damage using isogenic DNA repair‐deficient chicken DT40 cell lines , 2011, Environmental and molecular mutagenesis.

[28]  Ann Richard,et al.  Evaluation of high-throughput genotoxicity assays used in profiling the US EPA ToxCast chemicals. , 2009, Regulatory toxicology and pharmacology : RTP.

[29]  Chikahide Masutani,et al.  The XPV (xeroderma pigmentosum variant) gene encodes human DNA polymerase η , 1999, Nature.

[30]  Seth M. Cohen,et al.  Cisplatin: from DNA damage to cancer chemotherapy. , 2001, Progress in nucleic acid research and molecular biology.

[31]  M. Metzler,et al.  Induction of micronuclei, DNA strand breaks and HPRT mutations in cultured Chinese hamster V79 cells by the phytoestrogen coumoestrol. , 1997, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[32]  P. Burgers,et al.  The PCNA-RFC families of DNA clamps and clamp loaders. , 2004, Progress in nucleic acid research and molecular biology.

[33]  A. D’Andrea,et al.  Human ELG1 Regulates the Level of Ubiquitinated Proliferating Cell Nuclear Antigen (PCNA) through Its Interactions with PCNA and USP1* , 2010, The Journal of Biological Chemistry.

[34]  Adam Yasgar,et al.  Quantitative high-throughput screening: a titration-based approach that efficiently identifies biological activities in large chemical libraries. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[35]  F. Yoshimura,et al.  Genistein Induces Apoptosis in T Lymphoma Cells via Mitochondrial Damage , 2005, Nutrition and cancer.