DNA double-strand breaks by asbestos, silica, and titanium dioxide: possible biomarker of carcinogenic potential?

DNA double-strand breaks (DSBs) can result in cell death or genetic alterations when cells are subjected to radiation, exposure to toxins, or other environmental stresses. A complex DNA-damage-response pathway is activated to repair the damage, and the inability to repair these breaks can lead to carcinogenesis. One of the earliest responses to DNA DSBs is the phosphorylation of a histone, H2AX, at serine 139 (gamma-H2AX), which can be detected by a fluorescent antibody. A study was undertaken to compare the induction of DNA DSBs in normal (small airway epithelial) cells and cancer cells (A549) after exposure to asbestos (crocidolite), a proven carcinogen, silica, a suspected carcinogen, and titanium dioxide (TiO(2)), an inert particle recently reported to be carcinogenic in animals. The results indicate that crocidolite induced greater DNA DSBs than silica and TiO(2), regardless of cell type. DNA DSBs caused by crocidolite were higher in normal cells than in cancer cells. Silica and TiO(2) induced higher DNA DSBs in cancer cells than in normal cells. The production of reactive oxygen species was found to be highest in cells exposed to crocidolite, followed, in potency, by silica and TiO(2). The generation of reactive oxygen species was higher in normal cells than in cancer cells. Cell viability assay indicated that crocidolite caused the greatest cytotoxicity in both cell types. Apoptosis, measured by caspase 3/7 and poly (ADP-Ribose) polymerase activation, was highest in crocidolite-exposed cells, followed by TiO(2) and silica. The results of this study indicate that crocidolite has a greater carcinogenic potential than silica and TiO(2), judged by its ability to cause sustained genomic instability in normal lung cells.

[1]  Zbigniew Darzynkiewicz,et al.  Cytometric assessment of DNA damage in relation to cell cycle phase and apoptosis , 2005, Cell proliferation.

[2]  V. Castranova Signaling pathways controlling the production of inflammatory mediators in response to crystalline silica exposure: role of reactive oxygen/nitrogen species. , 2004, Free radical biology & medicine.

[3]  K. Jan,et al.  Ultrafine titanium dioxide particles in the absence of photoactivation can induce oxidative damage to human bronchial epithelial cells. , 2005, Toxicology.

[4]  M. Hedenborg Titanium dioxide induced chemiluminescence of human polymorphonuclear leukocytes , 1988, International archives of occupational and environmental health.

[5]  Xianglin Shi,et al.  Reactive oxygen species: their relation to pneumoconiosis and carcinogenesis. , 1998, Environmental health perspectives.

[6]  P. Olive,et al.  Expression of phosphorylated histone H2AX in cultured cell lines following exposure to X‐rays , 2003, International journal of radiation biology.

[7]  P. Olive,et al.  Expression of phosphorylated histone H2AX as a surrogate of cell killing by drugs that create DNA double-strand breaks. , 2003, Cancer research.

[8]  V. Vallyathan,et al.  Molecular Mechanisms of Asbestos- and Silica-Induced Lung Cancer , 2004 .

[9]  A. Kane,et al.  SV40 oncoproteins enhance asbestos-induced DNA double-strand breaks and abrogate senescence in murine mesothelial cells. , 2007, Cancer research.

[10]  A. Salinaro,et al.  Chemical oxidation and DNA damage catalysed by inorganic sunscreen ingredients , 1997, FEBS letters.

[11]  F. Fonnum,et al.  Evaluation of the probes 2',7'-dichlorofluorescin diacetate, luminol, and lucigenin as indicators of reactive species formation. , 2003, Biochemical pharmacology.

[12]  D. Kamp,et al.  Asbestos-Induced Pulmonary Toxicity: Role of DNA Damage and Apoptosis , 2003, Experimental biology and medicine.

[13]  B. Sarg,et al.  Hyperphosphorylation of histone H2A.X and dephosphorylation of histone H1 subtypes in the course of apoptosis , 2002, Cell Death and Differentiation.

[14]  Z. Darżynkiewicz,et al.  Cytometric assessment of histone H2AX phosphorylation: a reporter of DNA damage. , 2006, Methods in molecular biology.

[15]  X. Baur,et al.  Increased Incidence of DNA Double-strand Breaks and Anti-ds DNA Antibodies in Blood of Workers Occupationally Exposed to Asbestos , 1994, Human & experimental toxicology.

[16]  M. Fraga,et al.  Molecular analysis of a multistep lung cancer model induced by chronic inflammation reveals epigenetic regulation of p16 and activation of the DNA damage response pathway. , 2007, Neoplasia.

[17]  D. Nicholson,et al.  Analysis of titanium pigments in human lung tissue. , 1979, Scandinavian journal of work, environment & health.

[18]  Kyle Steenland,et al.  Silica, asbestos, man-made mineral fibers, and cancer , 1997, Cancer Causes & Control.

[19]  Priyadarshini Pande,et al.  Apoptosis induced by crocidolite asbestos in human lung epithelial cells involves inactivation of Akt and MAPK pathways , 2007, Apoptosis.

[20]  Xianglin Shi,et al.  Diseases caused by silica: mechanisms of injury and disease development. , 2002, International immunopharmacology.

[21]  T. Hei,et al.  New Insight into Intrachromosomal Deletions Induced by Chrysotile in the gpt delta Transgenic Mutation Assay , 2006, Environmental health perspectives.

[22]  Z. Darżynkiewicz,et al.  Induction of DNA double-strand breaks in A549 and normal human pulmonary epithelial cells by cigarette smoke is mediated by free radicals. , 2006, International journal of oncology.

[23]  Qamar Rahman,et al.  Evidence that ultrafine titanium dioxide induces micronuclei and apoptosis in Syrian hamster embryo fibroblasts. , 2002, Environmental health perspectives.

[24]  U Saffiotti,et al.  Silica radical-induced DNA damage and lipid peroxidation. , 1994, Environmental health perspectives.

[25]  R. Mason,et al.  Evidence for free radical formation during the oxidation of 2'-7'-dichlorofluorescin to the fluorescent dye 2'-7'-dichlorofluorescein by horseradish peroxidase: possible implications for oxidative stress measurements. , 1999, Free radical biology & medicine.

[26]  S. Reddy,et al.  Oxidants and signaling by mitogen-activated protein kinases in lung epithelium. , 2006, American journal of respiratory cell and molecular biology.

[27]  A. Ashworth,et al.  The roles of BRCA1 and BRCA2 and associated proteins in the maintenance of genomic stability , 2006, Oncogene.

[28]  J. Gamble,et al.  Silica, silicosis, and lung cancer: a response to a recent working group report. , 2000, Journal of occupational and environmental medicine.

[29]  Rachel L. Allen,et al.  Defying death after DNA damage , 2000, Nature.

[30]  D. Allison,et al.  Chromosomal Variations Within Aneuploid Cancer Lines , 2003, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[31]  K. Khanna,et al.  DNA double-strand breaks: signaling, repair and the cancer connection , 2001, Nature Genetics.

[32]  E. Rogakou,et al.  Megabase Chromatin Domains Involved in DNA Double-Strand Breaks in Vivo , 1999, The Journal of cell biology.

[33]  T. Hesterberg,et al.  In vitro toxicity of respirable-size particles of diatomaceous earth and crystalline silica compared with asbestos and titanium dioxide. , 1998, Journal of occupational and environmental medicine.

[34]  S. Weitzman,et al.  Asbestos causes apoptosis in alveolar epithelial cells: role of iron-induced free radicals. , 2001, The Journal of laboratory and clinical medicine.

[35]  R. Okayasu,et al.  Asbestos and DNA double strand breaks. , 1999, Cancer research.

[36]  J. Aten,et al.  Dynamics of DNA Double-Strand Breaks Revealed by Clustering of Damaged Chromosome Domains , 2004, Science.

[37]  S. Weitzman,et al.  The mitochondria-regulated death pathway mediates asbestos-induced alveolar epithelial cell apoptosis. , 2003, American journal of respiratory cell and molecular biology.

[38]  A. Churg,et al.  Pathology of Occupational Lung Disease , 1988 .

[39]  P. Borm,et al.  Chronic Inflammation and Tumor Formation in Rats After Intratracheal Instillation of High Doses of Coal Dusts, Titanium Dioxides, and Quartz , 2000, Inhalation toxicology.

[40]  W. Lutz,et al.  On the role of DNA double-strand breaks in toxicity and carcinogenesis. , 1997, Critical reviews in toxicology.

[41]  H. Ischiropoulos,et al.  Evaluation of the probe 2',7'-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress. , 1992, Chemical research in toxicology.

[42]  Z. Darżynkiewicz,et al.  Assessment of histone H2AX phosphorylation induced by DNA topoisomerase I and II inhibitors topotecan and mitoxantrone and by the DNA cross‐linking agent cisplatin , 2004, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[43]  V. Castranova,et al.  Oxidative and molecular interactions of multi-wall carbon nanotubes (MWCNT) in normal and malignant human mesothelial cells , 2008 .

[44]  E. Rogakou,et al.  DNA Double-stranded Breaks Induce Histone H2AX Phosphorylation on Serine 139* , 1998, The Journal of Biological Chemistry.