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Quantitative evaluation of the effects of human carcinogens and related chemicals on human foreskin fibroblasts

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Abstract

Ten compounds representative of diverse classes of chemicals were evaluated for their cytotoxicity and transforming ability to human skin fibroblasts in vitro. Only five of the ten compounds were highly cytotoxic in the 0-100 µg/ml range and their order of cytotoxicity was: 2,5-bis(1-aziridinyl)-3,6-bis(carboethoxyamino)-1,4-benzoquinone (AZQ) > cis platin > bis(chloromethyl)ether (BCME) > acrylonitrile > afatoxin BI (AFBI). The other five compounds, afatoxin B2 (AFB2), methylmethacrylate, 1-naphthylamine (1-NA), 2-naphthylamine (2-NA), and cyclophosphamide, exhibited less than 40% inhibition of colony formation even at 100 µg/ml of the compound (the maximum concentration of AFB2 used was 50 µg/ml due to its low solubility). Anchorage-independent growth of exposed cells in soft agar was used as a biological endpoint for the expression of chemical transformation. AFB1 had strong transforming ability, whereas AFB2 was a weak transforming agent. The transforming abilities of acrylonitrile, AZQ, BCME, cis-platin, methylmethacrylate and 2-NA ranged between those of AFBI and AFB2. 1-NA also induced the soft agar growth property in the treated cells even though this compound has not been shown to be carcinogenic. AFB1, AZQ, cisplatin, cyclophosphamide and 1-NA exhibited a dose dependent increase in soft agar growth frequency for at least three consecutive concentrations. The data suggest that anchorage-independent colony forming ability of exposed cells is a reliable marker to measure the carcinogenic potential of various hazardous chemicals.

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Abbreviations

AZQ:

2,5-bis(1-aziridinyl)-3,6-bis(carboethoxyamino)-1,4-benzoquinone

AFB:

aflatoxin B1

AFB2:

aflatoxin 132

AI:

anchorage independent

B[a]P:

benzo[a]pyrene

BCME:

bis(chloromethyl)ether; cis-platin, cis-diammine-dichloroplatinum

CM:

complete medium

E.D.50:

effective dose which produced 50% cytotoxicity

CP:

cyclophosphamide

HNF:

human neonatal foreskin

HPLC:

high pressure liquid chromatography

1-NA:

1-naphthylamine

2-NA:

2-naphthylamine

PDL:

population doubling

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Authors and Affiliations

  1. Department of Physiological Chemistry, The Ohio State University, 1645 Neil Avenue, 43210, Columbus, OH

    George E. Milo

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  1. Ponnamma Kurian
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  2. Stephen Nesnow
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  3. George E. Milo
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Kurian, P., Nesnow, S. & Milo, G.E. Quantitative evaluation of the effects of human carcinogens and related chemicals on human foreskin fibroblasts. Cell Biol Toxicol 6, 171–184 (1990). https://doi.org/10.1007/BF00249592

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