A Microassay for Measuring Cytochrome P450IA1 and Cytochrome P450IIB1 Activities in Intact Human and Rat Hepatocytes Cultured on 96-Well Plates

Donato, M. Teresa; Gómez‐Lechón, M. José; Castell, José V.

doi:10.1006/abio.1993.1381

Cited by 286 publications

(138 citation statements)

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“…This enzyme specifically dealkylates 7-pentoxyresorufin to the fluorescent compound, resorufin. 19 Stably transfected CYP2B1 cells were shown to display CYP2B1 enzymatic activity proportional to the cell number analysed. Typically one of the cell clones produces around 2 pmol/10 5 cells.…”

Section: Resultsmentioning

confidence: 99%

“…19 Before the day of measurement, different amounts of cells (2.8 × 10 6 , 5.5 × 10 5 , 5.5 × 10 4 ) were seeded into a 3-cm dish. On the day of testing the cells were washed with phosphate-buffered saline (PBS; Gibco) and overlayed with 500 l serum-free medium containing 15 m 7-pentoxy-resorufin (Sigma, Deisenhofen, Germany) and 10 m dicumarol (Sigma).…”

Section: Cytochrome P450 Enzymatic Assaymentioning

confidence: 99%

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Targeted chemotherapy by intratumour injection of encapsulated cells engineered to produce CYP2B1, an ifosfamide activating cytochrome P450

et al. 1998

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The prognosis of pancreatic adenocarcinoma is poor and agent ifosfamide to toxic metabolites. Administration of current treatment ineffective. A novel treatment strategy is ifosfamide to tumour-bearing mice that were recipients of described here using a mouse model system for pancreatic implanted encapsulated cells results in partial or even comcancer. Cells that have been genetically modified to plete tumour ablation. These results suggest that in situ express the cytochrome P450 2B1 enzyme are encapsuchemotherapy with genetically modified cells in an lated in cellulose sulphate and implanted into pre-estabimmunoprotected environment may prove useful for lished tumours derived from human pancreatic cells. application in man. Cytochrome P450 2B1 converts the chemotherapeutic

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Section: Resultsmentioning

confidence: 99%

Section: Cytochrome P450 Enzymatic Assaymentioning

confidence: 99%

Targeted chemotherapy by intratumour injection of encapsulated cells engineered to produce CYP2B1, an ifosfamide activating cytochrome P450

et al. 1998

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“…After incubation, EROD enzyme activity was analyzed following the methods of Donato et al (17) and Wang et al (18). Briefly, the culture medium was removed and was replaced with 100 µL fresh medium containing 10 µM 7-ethoxyresorufin (Sigma, E3763, United States) as substrate and 10 µM dicumarol (Sigma, M1390, United States).…”

Section: Methodsmentioning

confidence: 99%

Identification of Ah Receptor Agonists in Sediment of Meiliang Bay, Taihu Lake, China

Qiao

Chen

Zhang

et al. 2006

Environ. Sci. Technol.

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Deterioration of ecosystem and water quality in Taihu Lake, the third largest freshwater lake in China, is of great concern in China. However, most evaluations on water and sediment were carried out by chemical analysis, which could hardly gain information on the risk stressors affecting the system. Sediment samples from Meiliang Bay, Taihu Lake were assayed for AhR-mediated EROD induction using a rat hepatoma cell line (H4IIE). The known AhR agonists including PCDD/Fs, PCBs, and PAHs were analyzed by instruments. The cause-effect relationship between the observed EROD activity and chemical concentrations of AhR agonists and their dose-effect relationship were examined. Our results showed that sediment extracts could induce significant AhR effects, and the chemical-derived TCDD equivalents (TEQ cal ) were significantly correlated to bioassay-derived TCDD equivalents (TEQ bio ) (R ) 0.85, p < 0.01). Among different contributors, PAHs could account for 70-93% and PCDD/Fs could contribute less than 30% of TEQ cal . PCBs exhibited almost no contribution. Therefore, integration of EROD bioassay and chemical analysis provided an effective way for identifying the potential risk stressors. Under this circumstance, a quantitative doseeffect relationship between TEQ PAH and EROD activity could be established, suggesting that the observed AhR effect was mostly derived from PAHs. The implication is that the EROD assay and the identified risk stressors (PAHs) may be used in lieu of detailed environmental analysis to follow the progression of a remediation strategy.

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“…The cell culture and EROD bioassay were performed according to Donato et al (1993) with slight modifications as described in Schwirzer et al (1999). Briefly, cells were grown in Dulbecco's minimum essential medium supplemented with 10 % fetal calf serum and incubated at 37°C in a humidified 5 % CO 2 incubator.…”

Section: Micro-erod Bioassaymentioning

confidence: 99%

Aryl hydrocarbon receptor (AhR) inducers and estrogen receptor (ER) activities in surface sediments of Three Gorges Reservoir, China evaluated with in vitro cell bioassays

Wang

Bovee²,

et al. 2013

Environ Sci Pollut Res

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Two types of biological tests were employed for monitoring the toxicological profile of sediment cores in the Three Gorges Reservoir (TGR), China. In the present study, sediments collected in June 2010 from TGR were analyzed for estrogen receptor (ER)-and aryl hydrocarbon receptor (AhR)-mediated activities. The estrogenic activity was assessed using a rapid yeast estrogen bioassay, based on the expression of a green fluorescent reporter protein. Weak anti-estrogenic activity was detected in sediments from an area close to the dam of the reservoir, and weak estrogenic activities ranging from 0.3 to 1 ng 17β-estradiol (E2) equivalents (EQ) g −1 dry weight sediment (dw) were detected in sediments from the Wanzhou to Guojiaba areas. In the upstream areas Wanzhou and Wushan, sediments demonstrated additive effects in coadministration of 1 nM E2 in the yeast test system, while sediments from the downstream Badong and Guojiaba areas showed estrogenic activities which seemed to be more than additive (synergistic activity). There was an increasing tendency in estrogenic activity from upstream of TGR to downstream, while this tendency terminated and converted into anti-estrogenic activity in the area close to the dam. The AhR activity was detected employing rat hepatoma cell line (H4IIE). EROD activities were found homogenously distributed in sediments in TGR ranging from 200 to 311 pg 2,3,7,8-tetrachlorodibenzo-p -dioxin (TCDD) EQ g −1 dw for total AhR agonists and from 45 to 76 pg TCDD EQ g −1 dw for more persistent AhR agonists. The known AhR agonists polycyclic aromatic hydrocarbon, polychlorinated biphenyl, and PCDD/F only explained up to 8 % of the more persistent AhR agonist activity in the samples, which suggests that unidentified AhR-active compounds represented a great proportion of the TCDD EQ in sediments from TGR. These findings of estrogenic potential and dioxin-like activity in TGR sediments provide possible weight-of-evidence of potential ecotoxicological causes for the declines in fish populations which have been observed during the past decades in TGR.

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A Microassay for Measuring Cytochrome P450IA1 and Cytochrome P450IIB1 Activities in Intact Human and Rat Hepatocytes Cultured on 96-Well Plates

Cited by 286 publications

References 0 publications

Targeted chemotherapy by intratumour injection of encapsulated cells engineered to produce CYP2B1, an ifosfamide activating cytochrome P450

Targeted chemotherapy by intratumour injection of encapsulated cells engineered to produce CYP2B1, an ifosfamide activating cytochrome P450

Identification of Ah Receptor Agonists in Sediment of Meiliang Bay, Taihu Lake, China

Aryl hydrocarbon receptor (AhR) inducers and estrogen receptor (ER) activities in surface sediments of Three Gorges Reservoir, China evaluated with in vitro cell bioassays

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