An interlaboratory comparison of enhanced morphological transformation of Syrian hamster embryo cells cultured under conditions of reduced bicarbonate concentration and pH

LeBoeuf, Robert A.; Kerckaert, Gary A.; Poiley, J. A.; Raineri, R.

doi:10.1016/0165-1218(89)90136-5

Cited by 47 publications

(10 citation statements)

References 14 publications

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“…A description of the protocol for conduct of the reduced pH SHE cell transformation assay was originally published in 1989 (9). A more detailed description of this protocol is currently in press (7).…”

Section: Methodsmentioning

confidence: 99%

Use of the Syrian hamster embryo cell transformation assay for carcinogenicity prediction of chemicals currently being tested by the National Toxicology Program in rodent bioassays.

Kerckaert

Brauninger

LeBoeuf

et al. 1996

Environ Health Perspect

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The Syrian hamster embryo (SHE) cell transformation assay was used to predict the carcinogenicity of 26 chemicals currently being tested in the rodent bioassay by the National Toxicology Program as part of its program titled "Strategies for Predicting Chemical Carcinogenesis in Rodents." Of these 26 chemicals, 17 were found to be positive in the SHE cell transformation assay while 9 were negative. Carcinogenicity predictions were made for these chemicals, based upon the SHE cell transformation assay results. Our predictions will be compared with the rodent bioassay results as they become available. Environ Health Perspect

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Section: Methodsmentioning

confidence: 99%

Use of the Syrian hamster embryo cell transformation assay for carcinogenicity prediction of chemicals currently being tested by the National Toxicology Program in rodent bioassays.

Kerckaert

Brauninger

LeBoeuf

et al. 1996

Environ Health Perspect

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“…This effect of pH is not totally surprising, having been observed to play significant roles in the reproducibility of some standardized assays. 18,19 The maintenance of pH gradients between cellular components and the cytoplasm is known to be essential for correct functioning of the cellular endocytotic and secretory machinery, 20 and hence the ability to enhance uptake of chemical agents into tumor cells by manipulation of extracellular pH, 21 as well as allowing for alterations in the cell cycle of exposed cultures. 22 Therefore, changes in media pH could potentially alter the E 2 uptake and disposition within cellular compartments as well as possibly having an impact on the cell cycle, leading to changes in the expression of the reporter or other cellular proteins.…”

Section: Discussionmentioning

confidence: 99%

Optimization of an estrogen receptor-? transcriptional activation assay for testing a diverse spectrum of chemicals

Charles¹,

Gennings²,

Clemons³

et al. 2000

J. Appl. Toxicol.

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Estrogen receptor (ER) transactivation assays were initially designed to study endogenous mechanisms of steroid hormone action, but more recently have been used to assess industrial chemicals for potential estrogenic activity. Given the diverse spectrum of physicochemical properties of these chemicals, we examined the effects of pH (a weak organic and strong inorganic acid and base), hyperosmolality (NaCl, mannitol) and two different vehicles (DMSO, Triton X‐100) on responses to estradiol‐17β (E2) in an ER transactivation assay. MCF‐7 human breast cancer cells were transiently transfected with a chimeric estrogen receptor (Gal4‐HEG0) and a Gal4‐regulated luciferase reporter gene (17m5‐G‐Luc), treated with E2 under various test conditions, and then assessed for ER‐mediated luciferase activity. Maximal E2‐induced reporter activity was observed at pH 7.8 (pre‐incubation), but was markedly reduced at pH ⩽7.5 or ⩾8.0 (P < 0.001), even though there was no evidence of cytotoxicity. Hyperosmolality induced by addition of mannitol (⩾25 mM) resulted in significant decreases in overall assay responsiveness, whereas NaCl (⩾80 mM) decreased the sensitivity of the assay by increasing the no‐observed‐effect level for E2 compared to control cultures (330 mOsm). The maximal DMSO concentration that resulted in consistently high E2‐induced reporter activity was 0.1%, whereas concentrations of Triton X‐100 above 1 ppm inhibited E2‐induced reporter responses and were cytotoxic above 10 ppm. These results indicate that various physicochemical factors have the potential to confound assay data if not kept within predefined operational limits. Copyright © 2000 John Wiley & Sons, Ltd.

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“…The assay which we used has been already described in detail (Pienta et al, 1977;Jones et al, 1988;LeBoeuf et al, 1989). We modified the method in two steps (a) we used UV-irradiation of feeder cells (40 J/m 2) instead of X-irradiation (5000 R), (b) the ratio of feeder and target cells was 8 × 104 to 3 × 102.…”

Section: Morphological Transformation Of She Cellsmentioning

confidence: 99%

An evaluation of three pesticides: Piritione, supercypermethrin and metolachlor in transformation bioassays of BHK21 and hamster embryo cells

et al. 1992

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In a study of potential carcinogenicity of pesticides, Piritione, metolachlor (in the form of Dual and VUCHT 524) and Supercypermethrin (in the form of Supercypermethrin EC and Supercypermethrin TP) were assayed for induction of anchorage independent growth of BHK21 cells and morphological transformation of Syrian hamster embryo cells. The activity of these substances in both transformation assays was compared to the activity of the direct-acting ultimate carcinogen N-methyl-N-nitrosourea. In comparison to the very strong transforming activity of N-methyl-N-nitrosourea all pesticides tested with or without S9 fraction manifested a very weak, weak, medium or strong effect. The ability to induce anchorage independent growth was graded as follows: Dual < Supercypermethrin EC < Supercypermethrin TP < or = Piritione < VUCHT 524. Results of Syrian hamster embryo cell transformation assay were very similar to the BKH21 transformation assay. VUCHT 524 strongly induced transformation whereas Dual was inactive. Piritione and Supecypermethrin EC and Supercypermethrin TP elicited a slight but significant positive response.

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An interlaboratory comparison of enhanced morphological transformation of Syrian hamster embryo cells cultured under conditions of reduced bicarbonate concentration and pH

Cited by 47 publications

References 14 publications

Use of the Syrian hamster embryo cell transformation assay for carcinogenicity prediction of chemicals currently being tested by the National Toxicology Program in rodent bioassays.

Use of the Syrian hamster embryo cell transformation assay for carcinogenicity prediction of chemicals currently being tested by the National Toxicology Program in rodent bioassays.

Optimization of an estrogen receptor-? transcriptional activation assay for testing a diverse spectrum of chemicals

An evaluation of three pesticides: Piritione, supercypermethrin and metolachlor in transformation bioassays of BHK21 and hamster embryo cells

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