Quantitative Immunohistochemical Analysis of 4-Aminobiphenyl-DNA in Cultured Cells and Mice: Comparison to Gas Chromatography/Mass Spectroscopy Analysis

Al-Atrash, Jenan; Zhang, Yujing; Lin, Dong; Kadlubar, Fred F.; Santella, Regina M.

doi:10.1021/tx00047a015

Cited by 28 publications

(17 citation statements)

References 10 publications

(17 reference statements)

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“…The extra peak at a retention time of 22.029 min in the acid hydrolysate of modified DNA is characteristic of dG-C8-4-ABP adduct, a known marker for DNA damaged by 4-ABP and N -OH-AABP [33]. The identification of dG-C8-4-ABP adduct is in agreement with the published reports on DNA-adducts with arylamines in experimental animals [15]–[17]. Similar DNA-adducts have been reported in the biopsy samples of human urinary bladder [18], [19].…”

Section: Resultssupporting

confidence: 88%

“…The primary step is N -oxidation of arylamines and arylacetamides by specific cytochrome P 450 (CYP1A2) in hepatic tissues [10], followed by conjugation of the N -hydroxyl function with acetate, sulfate, or glucuronate [11]–[14]. The activated electrophilic derivatives of 4-ABP exert their genotoxic effect through interaction with DNA forming adducts that have been reported for a role in bladder carcinogenesis both in experimental animals [15]–[17] and humans [18], [19]. DNA adducts have been also reported upon exposure of human bladder cells to N -OH-ABP, N -OH-AABP and N -OAc-AABP [20]–[22].…”

Section: Introductionmentioning

confidence: 99%

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Genotoxic Effect of N-Hydroxy-4-Acetylaminobiphenyl on Human DNA: Implications in Bladder Cancer

et al. 2013

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BackgroundThe interaction of environmental chemicals and their metabolites with biological macromolecules can result in cytotoxic and genotoxic effects. 4-Aminobiphenyl (4-ABP) and several other related arylamines have been shown to be causally involved in the induction of human urinary bladder cancers. The genotoxic and the carcinogenic effects of 4-ABP are exhibited only when it is metabolically converted to a reactive electrophile, the aryl nitrenium ions, which subsequently binds to DNA and induce lesions. Although several studies have reported the formation of 4-ABP-DNA adducts, no extensive work has been done to investigate the immunogenicity of 4-ABP-modified DNA and its possible involvement in the generation of antibodies in bladder cancer patients.Methodology/Principal FindingsHuman DNA was modified by N-hydroxy-4-acetylaminobiphenyl (N-OH-AABP), a reactive metabolite of 4-ABP. Structural perturbations in the N-OH-AABP modified DNA were assessed by ultraviolet, fluorescence, and circular dichroic spectroscopy as well as by agarose gel electrophoresis. Genotoxicity of N-OH-AABP modified DNA was ascertained by comet assay. High performance liquid chromatography (HPLC) analysis of native and modified DNA samples confirmed the formation of N-(deoxyguanosine-8-yl)-4-aminobiphenyl (dG-C8-4ABP) in the N-OH-AABP damaged DNA. The experimentally induced antibodies against N-OH-AABP-modified DNA exhibited much better recognition of the DNA isolated from bladder cancer patients as compared to the DNA obtained from healthy individuals in competitive binding ELISA.Conclusions/SignificanceThis work shows epitope sharing between the DNA isolated from bladder cancer patients and the N-OH-AABP-modified DNA implicating the role of 4-ABP metabolites in the DNA damage and neo-antigenic epitope generation that could lead to the induction of antibodies in bladder cancer patients.

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Section: Resultssupporting

confidence: 88%

Section: Introductionmentioning

confidence: 99%

Genotoxic Effect of N-Hydroxy-4-Acetylaminobiphenyl on Human DNA: Implications in Bladder Cancer

et al. 2013

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“…The 4C11 antibody is highly specific for the 4-ABP-DNA adduct and, at the highest concentration tested, does not recognize the DNA adducts of several other aromatic amines, including 1-aminopyrene, 8-nitro-1-aminopyrene, and 6-nitro-1-aminopyrene (27). The Immunodot blot assay was conducted as described earlier with some modifications (30).…”

Section: Immunodot Blot Assaymentioning

confidence: 99%

“…injections of 4-ABP for 6 weeks, followed by a 6-week recovery period. We have used an Immunodot blot assay with a specific antibody raised against the 4-ABP-DNA adduct (27) to evaluate DNA damage and repair in bladder and various nontarget organs of 4-ABP-treated mice. In addition, we have used the cII mutation detection assay (28) to assess the organ specificity of cII mutations, and DNA sequencing analysis to establish the type and frequency distribution of induced mutations in 4-ABP-treated animals.…”

Section: Introductionmentioning

confidence: 99%

Organ Specificity of the Bladder Carcinogen 4-Aminobiphenyl in Inducing DNA Damage and Mutation in Mice

Yoon

Kim

Tommasi

et al. 2012

Cancer Prevention Research

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Aromatic amines are a widespread class of environmental contaminants present in various occupational settings and tobacco smoke. Exposure to aromatic amines is a major risk factor for bladder cancer development. The etiologic involvement of aromatic amines in the genesis of bladder cancer is attributable to their ability to form DNA adducts, which upon eluding repair and causing mispairing during replication, may initiate mutagenesis. We have investigated the induction of DNA adducts in relation to mutagenesis in bladder and various nontarget organs of transgenic Big Blue mice treated weekly (i.p.) with a representative aromatic amine compound, 4-aminobiphenyl (4-ABP), for six weeks, followed by a six-week recovery period. We show an organ-specificity of 4-ABP in inducing repair-resistant DNA adducts in bladder, kidney, and liver of carcinogen-treated animals, which accords with the bioactivation pathway of this chemical in the respective organs. In confirmation, we show a predominant and sustained mutagenic effect of 4-ABP in bladder, and much weaker but significant mutagenicity of 4-ABP in the kidney and liver of carcinogentreated mice, as reflected by the elevation of background cII mutant frequency in the respective organs. The spectrum of mutations produced in bladder of 4-ABP-treated mice matches the known mutagenic properties of 4-ABP-DNA adducts, as verified by the preponderance of induced mutations occurring at G:C base pairs (82.9%), with the vast majority being G:C!T:A transversions (47.1%). Our data support a possible etiologic role of 4-ABP in bladder carcinogenesis and provide a mechanistic view on how DNA adduct-driven mutagenesis, specifically targeted to bladder urothelium, may account for organ-specific tumorigenicity of this chemical. Cancer Prev Res; 5(2); 299-308. Ó2011 AACR.

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“…In order to denature DNA, the samples were incubated with 4N HCl for 10 min and with 50mM TRIS Base for 5 min at room temperature, which is an essential step for adduct evaluation. 18 After washing with PBS, the samples were incubated with 0.3% H 2 O 2 in methyl alcohol at room temperature for 30 min to quench endogenous peroxidase activity. Nonspecific binding was blocked with 1.5% normal horse serum and incubation with the monoclonal antibody 1F7 12 diluted 1:50 in horse serum was performed overnight at þ 41C.…”

Section: Immunoperoxidase Detection Of 8-ohdgmentioning

confidence: 99%

Expression of the CDK inhibitor p27kip1 and oxidative DNA damage in non-neoplastic and neoplastic vulvar epithelial lesions

et al. 2006

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Vulvar cancer represents an important medical problem worldwide whose incidence is increasing at an alarming rate in young females. Several factors have been linked to vulvar cancer development, but its exact pathogenesis remains to be determined. Vulvar tumorigenesis proceeds through intermediate dysplastic lesions, known as vulvar intraepithelial neoplasias, frequently associated with non-neoplastic epithelial disorders of the vulva, such as lichen sclerosus and squamous cell hyperplasia. In this study, the expression of the CDK inhibitor p27 Kip1 and the extent of endogenous oxidative DNA damage were evaluated in vulvar specimens, including normal tissues, lichen sclerosus, squamous cell hyperplasia, vulvar intraepithelial neoplasias and invasive squamous cell carcinomas. We found that p27 Kip1 was constantly expressed in normal vulvar epithelium cells while a progressive significant reduction in the percentage of p27 Kip1 -positive cells was observed in vulvar intraepithelial neoplasias (77%) and in invasive carcinomas (64%). Mean percentage of positive cells in invasive carcinomas, but not in vulvar intraepithelial neoplasias, was also significantly lower than squamous cell hyperplasia lesions (78%) while lichen sclerosus displayed a percentage of positive cells (45%) significantly lower than both vulvar intraepithelial neoplasias and invasive carcinomas. 8-hydroxydeoxyguanosine (8-OHdG) is considered a sensitive biomarker for oxidative stress. We observed a progressive significant increase in the levels of 8-OHdG and in the percentage of positive cells from normal vulvar epithelium to vulvar intraepithelial neoplasias (25%) and to invasive carcinomas (64%). Squamous cell hyperplasia displayed an intermediate percentage of positive cells comparable to vulvar intraepithelial neoplasias 2 but significantly higher than vulvar intraepithelial neoplasias 1 and lower than invasive carcinomas. Lichen sclerosus staining was significantly lower than carcinomas but higher than vulvar intraepithelial neoplasias and squamous cell hyperplasia. These results demonstrate that expression of p27 Kip1 is downregulated while oxidative DNA damage increases from early non-neoplastic epithelial alterations through vulvar intraepithelial neoplasias to invasive vulvar carcinomas. Thus, both parameters might play an important role in the development of this cancer and their study might contribute to our understanding of human vulvar carcinogenesis. Keywords: vulvar cancer; tumorigenesis; cell cycle; oxidative DNA damage Squamous cell carcinoma of the vulva accounts for 4% of all gynecologic malignancies and although its incidence rates in older women have remained relatively stable over the past few decades, diagnosed cases of vulvar cancer are increasing at an alarming rate in young females worldwide. 1 The type of vulvar squamous cell carcinoma usually found in younger women appears to be associated with human papillomavirus (HPV) infection. Indeed, specific genital HPV types, in particular HPV 16, are strongly implicated in the ...

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Quantitative Immunohistochemical Analysis of 4-Aminobiphenyl-DNA in Cultured Cells and Mice: Comparison to Gas Chromatography/Mass Spectroscopy Analysis

Cited by 28 publications

References 10 publications

Genotoxic Effect of N-Hydroxy-4-Acetylaminobiphenyl on Human DNA: Implications in Bladder Cancer

Genotoxic Effect of N-Hydroxy-4-Acetylaminobiphenyl on Human DNA: Implications in Bladder Cancer

Organ Specificity of the Bladder Carcinogen 4-Aminobiphenyl in Inducing DNA Damage and Mutation in Mice

Expression of the CDK inhibitor p27kip1 and oxidative DNA damage in non-neoplastic and neoplastic vulvar epithelial lesions

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