An in Vitro Reporter Gene Assay Method Incorporating Metabolic Activation with Human and Rat S9 or Liver Microsomes

Sumida, Kayo; Ooe, Norihisa; Nagahori, Hirohisa; Saito, Koichi; Isobe, Naohiko; Kaneko, Hideo; Nakatsuka, Iwao

doi:10.1006/bbrc.2000.4071

Cited by 37 publications

(27 citation statements)

References 19 publications

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“…Sumithrin showed weak estrogenic activity based on a maximum MCF-7 BUS cell proliferation that was equivalent to 70% of that of 17β-estradiol. These results are in partial agreement with those of previous reports in which pyrethroid insecticides did not show any estrogenic activity in MCF-7 cell proliferation assay [11], and that certain pyrethroid insecticides (fenvalerate, d-phenothrin, permethrin, and cypermethrin) did not exhibit estrogenicity in a reporte r g ene assay w ith or without liver microsomes or S9 mix [12]. In contrast, Go et al [10] found that fenvalerate, d-allethrin, and sumithrin, at concentrations of 10 -5 M, induced MCF-7 cell proliferation at a level comparable to the effect of 17β -estradiol.…”

Section: Discussionsupporting

confidence: 92%

“…On the other hand, certain pyrethroid insecticides did not show any estrogenic or antiestrogenic activity in vitro [11]. Moreover, four pyrethroid insecticides (permethrin, cypermethrin, fenvalerate, and phenothrin) did not induce estrogenic activity in an in vitro luciferase reporter gene assay using the metabolic activation system [12]. These results were confirmed with a uterotrophic assay in which none of the pyrethroid insecticides tested (esfenvalerate, fenvalerate, and permethrin) exhibited any estrogenic activity [13].…”

mentioning

confidence: 74%

“…These compounds are suspected EDs [1]. Recently, some pyrethroid insecticides were evaluated for their estrogenic or anti-estrogenic activities using in vitro combination assays, such as cell proliferation and estrogensensitive pS2 expression assays [9][10][11][12]. However, the results of these studies are controversial.…”

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confidence: 99%

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Assessing Estrogenic Activity of Pyrethroid Insecticides Using In Vitro Combination Assays

et al. 2004

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Abstract. Pyrethroid insecticides are among the most commonly used classes of insecticides worldwide, but their endocrine disrupting activities remain unclear. Therefore, in the present study, we examined the estrogenic activities of pyrethroid insecticides in E-screen and competition binding assays. In addition, we measured estrogen receptor (ER) protein and pS2 mRNA levels in human breast cancer cells (MCF-7 BUS) to clarify the mechanism of their estrogenicity. Seven pyrethroid insecticides (bioallethrine, cypermethrin, deltamethrin, fenvalerate, permethrin, sumithrin, and tetramethrin) were tested because of their worldwide usage. In addition, 17β-estradiol was tested as a positive control. As expected, 17β-estradiol significantly increased MCF-7 BUS cell proliferation at concentrations of 10 -11 M and above. Of the pyrethroid insecticides tested, only sumithrin increased MCF-7 BUS cell proliferation in a dose-dependent manner; the maximum induction of cell proliferation was observed at a dose of 10 -5 M. In the anti-estrogenic activity test, bioallethrin, fenvalerate, and permethrin significantly inhibited 17β-estradiol-induced MCF-7 BUS cell proliferation at 10 -6 M, a concentration comparable to the effective dose (10 -9 M) of ICI 182,780, a pure ER antagonist. However, none of the pyrethroid insecticides competitively inhibited the binding of [ 3 H]estradiol to rat uterus ERs in competition binding assays. Both 17β-estradiol (10 -10 M) and sumithrin (10 -5 M) decreased the levels of cytosolic ERα and ERβ protein expression significantly as compared with the vehicle control. In addition, 17β-estradiol (10 -10 M) increased pS2 mRNA expression markedly, and sumithrin significantly increased pS2 mRNA levels in a dose-dependent manner. The other six compounds tested in the present study did not affect ER protein levels or pS2 mRNA levels. These results suggest that certain pyrethroid insecticides may be considered to be estrogen-like chemicals that act through pathways other than direct ER binding, and may function as endocrine modulators in both wildlife and humans.

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Section: Discussionsupporting

confidence: 92%

mentioning

confidence: 74%

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Assessing Estrogenic Activity of Pyrethroid Insecticides Using In Vitro Combination Assays

et al. 2004

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“…Various pesticides might be converted to active estrogens by microsomal oxidase systems, though Sumida et al showed that perimethrine was not metabolically activated. 79) p-Hydroxybenzophenone, which is formed from benzophenone, an antifungal agent, in rat hepatocytes, is also estrogenic. 80,81) It is also reported that anethole, a flavor agent, is not estrogenic, but 4-hydroxy-1-propenylbenzene, the desmethylated metabolite of anethole, exhibited estrogenic activity.…”

Section: Activation Of Other Proestrogens In Environments By the Cytomentioning

confidence: 99%

Metabolic Activation of Proestrogens in the Environment by Cytochrome P450 System

Kawa

Sugihara

Sanoh

et al. 2008

JOURNAL OF HEALTH SCIENCE

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Liver microsome-mediated activation of proestrogens in the environment is reviewed here. Proestrogens such as methoxychlor, trans-stilbene, diphenyl, diphenylmethane, 2,2-diphenylpropane, benzo[a]pyrene, benzophenone, 2-nitrofluorene (NF), chalcone, trans-4-phenyl-3-buten-2-one and styrene oligomers are negative in in vitro estrogen screening tests. However, those proestrogens exhibit estrogenic activity after metabolic activation by the microsomal cytochrome P450 system. In these cases, hydroxylated derivatives of the compounds are formed as major metabolites, and these metabolites exhibit significant estrogenic activities. Thus, the estrogenic activities of proestrogenic compounds are a consequence of metabolism of the parent compounds. Various candidates for proestrogens among medicines and insecticides are also discussed.

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“…Methods including receptor binding assays, [2][3][4] a MCF-7 cell proliferation assay (E-screen) 5) and reporter gene assays 6,7) on animal cell lines have recently been developed as in vitro screening techniques to detect endocrine disruptors. A yeast twohybrid assay recently developed by Nishikawa et al 8) has attracted attention because it allows prediction of endocrine disruptors in a relatively short period of time.…”

Section: Introductionmentioning

confidence: 99%

Problems with Methods used to Screen Estrogenic Chemicals by Yeast Two-Hybrid Assays.

Nakano

Nagao

Kobayashi

et al. 2002

JOURNAL OF HEALTH SCIENCE

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We developed a yeast two-hybrid assay using a human-type estrogen receptor (hERα) to screen test chemicals reporting weak estrogenic activity at high concentrations. As a result of exposure to several test chemicals, viable yeast cells showed a decrease in number compared with a control. It was shown that this phenomenon was due to fungicidal effects of the test chemicals within the yeast cell or to inhibition of yeast cell proliferation. Therefore, in performing a yeast twohybrid assay, it is necessary to monitor yeast cell proliferation by methods such as the measurement of the number of viable cells, and it is necessary to screen test chemicals within a concentration range where these chemicals show neither fungicidal effects towards the yeast cell nor inhibition of yeast cell proliferation.

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An in Vitro Reporter Gene Assay Method Incorporating Metabolic Activation with Human and Rat S9 or Liver Microsomes

Cited by 37 publications

References 19 publications

Assessing Estrogenic Activity of Pyrethroid Insecticides Using In Vitro Combination Assays

Assessing Estrogenic Activity of Pyrethroid Insecticides Using In Vitro Combination Assays

Metabolic Activation of Proestrogens in the Environment by Cytochrome P450 System

Problems with Methods used to Screen Estrogenic Chemicals by Yeast Two-Hybrid Assays.

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