A stable and sensitive testing system for potential carcinogens based on DNA damage-induced gene expression in human HepG2 cell

Zhang, Rong; Niu, Yujie; Du, Hongwu; Cao, Xin; Shi, Dan; Qiaoling, Hao; Zhou, Yikai

doi:10.1016/j.tiv.2008.10.006

Cited by 14 publications

(8 citation statements)

References 31 publications

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“…The LOEC for BaP was at 0.13 μg/mL (0.5 μM), and at the highest tested concentration 1.26 μg/mL (5 μM) the relative EGFP induction ratio was 8.54 after 24 h exposure. HepG2 cells transfected with GADD153 fused to luciferase were significantly more sensitive for BaP genotoxicity detection; the LOEC was 0.0025 μg/mL (10 nM) 29. The authors ascribed high sensitivity of their assay compared to other reporter systems to the sensitivity of luciferase, which seems to be higher than that of EGFP 29.…”

Section: Discussionmentioning

confidence: 97%

“…The assay is commercially available as GreenScreen HC assay provided by Gentronics Ltd (UK). Recently Zhang et al 29. developed a stably transfected HepG2 cell line containing GADD153 promoter regions coupled to the luciferase reporter gene.…”

Section: Discussionmentioning

confidence: 99%

“…HepG2 cells transfected with GADD153 fused to luciferase were significantly more sensitive for BaP genotoxicity detection; the LOEC was 0.0025 μg/mL (10 nM) 29. The authors ascribed high sensitivity of their assay compared to other reporter systems to the sensitivity of luciferase, which seems to be higher than that of EGFP 29. In MCF-7 cells transfected with p532R coupled to the luciferase reporter gene, the LOEC for BaP was 0.26 μg/mL when tested without metabolic activation and 0.12 μg/mL in the presence of metabolic activation 24.…”

Section: Discussionmentioning

confidence: 99%

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Development of human cell biosensor system for genotoxicity detection based on DNA damage-induced gene expression

Žager¹,

Čemažar²,

Hreljac³

et al. 2010

Radiology and Oncology

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BackgroundHuman exposure to genotoxic agents in the environment and everyday life represents a serious health threat. Fast and reliable assessment of genotoxicity of chemicals is of main importance in the fields of new chemicals and drug development as well as in environmental monitoring. The tumor suppressor gene p21, the major downstream target gene of activated p53 which is responsible for cell cycle arrest following DNA damage, has been shown to be specifically up-regulated by genotoxic carcinogens. The aim of our study was to develop a human cell-based biosensor system for simple and fast detection of genotoxic agents.MethodsMetabolically active HepG2 human hepatoma cells were transfected with plasmid encoding Enhanced Green Fluorescent Protein (EGFP) under the control of the p21 promoter (p21HepG2GFP). DNA damage was induced by genotoxic agents with known mechanisms of action. The increase in fluorescence intensity, due to p21 mediated EGFP expression, was measured with a fluorescence microplate reader. The viability of treated cells was determined by the colorimetric MTS assay.ResultsThe directly acting alkylating agent methylmethane sulphonate (MMS) showed significant increase in EGFP production after 48 h at 20 μg/mL. The indirectly acting carcinogen benzo(a)pyren (BaP) and the cross-linking agent cisplatin (CisPt) induced a dose- dependent increase in EGFP fluorescence, which was already significant at concentrations 0.13 μg/mL and 0.41 μg/mL, respectively. Vinblastine (VLB), a spindle poison that does not induce direct DNA damage, induced only a small increase in EGFP fluorescence intensity after 24 h at the lowest concentration (0.1 μg/mL), while exposure to higher concentrations was associated with significantly reduced cell viability.ConclusionsThe results of our study demonstrated that this novel assay based on the stably transformed cell line p21HepG2GFP can be used as a fast and simple biosensor system for detection of genetic damage caused by chemical agents.

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Section: Discussionmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Development of human cell biosensor system for genotoxicity detection based on DNA damage-induced gene expression

Žager¹,

Čemažar²,

Hreljac³

et al. 2010

Radiology and Oncology

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“…After DDT absorption, it is distributed to all organs and a portion will be stored in fatty tissues, especially if the exposure was massive [141][142][143][144][145][146] . Many insecticides, including DDT, were reported to be responsible for leading the development of HCC [147][148][149][150][151][152] .…”

Section: Pesticidesmentioning

confidence: 99%

Hepatocellular carcinoma and the risk of occupational exposure

Rapisarda

Loreto

Malaguarnera

et al. 2016

WJH

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Hepatocellular carcinoma (HCC) is the most common type of liver cancer. The main risk factors for HCC are alcoholism, hepatitis B virus, hepatitis C virus, nonalcoholic steatohepatitis, obesity, type 2 diabetes, cirrhosis, aflatoxin, hemochromatosis, Wilson's disease and hemophilia. Occupational exposure to chemicals is another risk factor for HCC. Often the relationship between occupational risk and HCC is unclear and the reports are fragmented and inconsistent. This review aims to summarize the current knowledge regarding the association of infective and non-infective occupational risk exposure and HCC in order to encourage further research and draw attention to this global occupational public health problem. Core tip: Hepatocellular carcinoma (HCC) is the fifth most common human cancer. This review summarizes current knowledge regarding the occupational risk factors of HCC. In particular, we underline not only the infective but also non-infective occupational risk exposure, including chemical agents and toxic metabolites which are a major cause of liver damage.

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“…TFF1 is suspected of playing a direct role in mucus polymerization [48] and mucus viscosity [53], while CA2 is a key enzyme for reducing acidity through bicarbonate buffering [44]. Given that gastric acid can cause double-stranded DNA damage within exposed BE tissue [54], a breakdown in the mucosal defence system could contribute to the frequent chromosomal damage seen in EAC [24], [55], [56], [57]. More research is required in this regard.…”

Section: Resultsmentioning

confidence: 99%

Whole Genome Expression Array Profiling Highlights Differences in Mucosal Defense Genes in Barrett's Esophagus and Esophageal Adenocarcinoma

et al. 2011

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Esophageal adenocarcinoma (EAC) has become a major concern in Western countries due to rapid rises in incidence coupled with very poor survival rates. One of the key risk factors for the development of this cancer is the presence of Barrett's esophagus (BE), which is believed to form in response to repeated gastro-esophageal reflux. In this study we performed comparative, genome-wide expression profiling (using Illumina whole-genome Beadarrays) on total RNA extracted from esophageal biopsy tissues from individuals with EAC, BE (in the absence of EAC) and those with normal squamous epithelium. We combined these data with publically accessible raw data from three similar studies to investigate key gene and ontology differences between these three tissue states. The results support the deduction that BE is a tissue with enhanced glycoprotein synthesis machinery (DPP4, ATP2A3, AGR2) designed to provide strong mucosal defenses aimed at resisting gastro-esophageal reflux. EAC exhibits the enhanced extracellular matrix remodeling (collagens, IGFBP7, PLAU) effects expected in an aggressive form of cancer, as well as evidence of reduced expression of genes associated with mucosal (MUC6, CA2, TFF1) and xenobiotic (AKR1C2, AKR1B10) defenses. When our results are compared to previous whole-genome expression profiling studies keratin, mucin, annexin and trefoil factor gene groups are the most frequently represented differentially expressed gene families. Eleven genes identified here are also represented in at least 3 other profiling studies. We used these genes to discriminate between squamous epithelium, BE and EAC within the two largest cohorts using a support vector machine leave one out cross validation (LOOCV) analysis. While this method was satisfactory for discriminating squamous epithelium and BE, it demonstrates the need for more detailed investigations into profiling changes between BE and EAC.

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A stable and sensitive testing system for potential carcinogens based on DNA damage-induced gene expression in human HepG2 cell

Cited by 14 publications

References 31 publications

Development of human cell biosensor system for genotoxicity detection based on DNA damage-induced gene expression

Development of human cell biosensor system for genotoxicity detection based on DNA damage-induced gene expression

Hepatocellular carcinoma and the risk of occupational exposure

Whole Genome Expression Array Profiling Highlights Differences in Mucosal Defense Genes in Barrett's Esophagus and Esophageal Adenocarcinoma

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