Growth inhibition and transformation of a human fetal tracheal epithelial cell line by long-term exposure to diethylnitrosamine |
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Authors: | Emura Makito; Mohr Ulrich; Kakunaga Takeo; Hilfrich Jorn |
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Institution: | 1Institut für Experimentelle Pathologie, Zentrum für Pathologie und Rechtsmedizin Konstanty-Gutschow-Strasse 8, D-3000 Hannover 61, FRG
2Cell Genetics Section, Laboratory of Molecular Carcinogenesis, National Cancer Institute Bethesda, MD 20205, USA
3Abteilung f r Geburtshilfe und Gyn kologische Onkologie, Zentrum der Frauenheilkunde, Medizinische Hochschule Hannover Konstanty-Gutschow-Strasse 8, D-3000 Hannover 61, FRG |
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Abstract: | In order to obtain more information on the in vitro transformationof human cells, a human fetal tracheal epithelial cell line(FHET16/5) was exposed for a long time to diethylnitrosamine(DEN). In 20 passages, this cell line (diploid, male) maintainedstrong immunohistochemical reactivity for carcino-embryonnicantigen and wool merokeratin; it was negative for vimentin.The cells contained PAS-positive mucous substances and ultrastructurallywere found to have desmosomelike attachments. Treatment of thecells was with 0.3% dimethyl sulphoxide (DMSO), or DMSO with150, 450, 1000 or 2000 µg/ml of DEN. It was started atthe ninth passage and continued for six passages over 9 weeksfor the control (DMSO) and the three lowest control doses ofDEN, and for three passages over 9 weeks for the 2000 µg/mlDEN group. Cells grown for 13 days after the end of treatmentwere plated in soft agar and injected subcutaneously in nudemice. The frequency of anchorage-independent colonies grownin soft agar was directly related to DEN dose. Colony-formingefficiency, as an expression of toxic effect, was also dosedependent. Autoradiographically detected unscheduled DNA synthesisindicated an association between anchorage-independent transformationand DNA alterations induced by DEN. Cells injected into nudemice did not produce tumours during a 6-month period, but invasivenesswas observed when cells from the 2000 µg/ml DEN groupwere transplanted on the dermis of cultured chick embryo skin.The results indicate that DEN causes anchorage-independent transformationaccompanied by unscheduled DNA synthesis in a fetal human trachealepithelial cell line. |
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