SV40-transfected human melanocyte sensitivity to growth inhibition by the phorbol ester 12-O-tetradecanoylphorbol-13-acetate

Normal human melanocytes, which require the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) for growth in culture, were transfected with a SV40 T-antigen-containing plasmid by using the technique of electroporation, and were scored for colonies of morphologically altered cells. The frequenc...

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Bibliographic Details
Published inCancer research (Chicago, Ill.) Vol. 49; no. 13; pp. 3650 - 3655
Main Authors MELBER, K, GEFENG ZHU, DIAMOND, L
Format Journal Article
LanguageEnglish
Published Philadelphia, PA American Association for Cancer Research 01.07.1989
Subjects
Antigens, Polyomavirus Transforming - analysis
Biological and medical sciences
Cell Division - drug effects
Cell physiology
Cell transformation and carcinogenesis. Action of oncogenes and antioncogenes
Cell Transformation, Viral
Cells, Cultured
Electricity
Fluorescent Antibody Technique
Fundamental and applied biological sciences. Psychology
Humans
In Vitro Techniques
Karyotyping
Melanocytes - cytology
Molecular and cellular biology
Simian virus 40
Tetradecanoylphorbol Acetate - pharmacology
Transfection
Human
Cell proliferation
Cell culture
Transformed cell
Polyomavirus
Melanocyte
Electroporation
Phase contrast microscopy
Papovaviridae
Virus
Regulation(control)
Transfection
Immunofluorescence
SV40 virus
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Summary:Normal human melanocytes, which require the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) for growth in culture, were transfected with a SV40 T-antigen-containing plasmid by using the technique of electroporation, and were scored for colonies of morphologically altered cells. The frequency of transformed colonies was higher when selection was done in the absence rather than the presence of TPA in the medium. Three cell lines derived from transformed colonies were characterized. All show an enhanced growth rate compared to parental cells, anchorage independence, loss of dependence on medium supplements for growth, chromosomal abnormalities, an extended life span, and growth inhibition by TPA. They express nerve growth factor receptor and Mr 97,000 protein, melanotransferrin, two antigens usually associated with melanocytic cells, but the transformed cells are not pigmented. The three cell lines underwent crisis at about passage 10 posttransfection; one cell line recovered and appears to have unlimited growth potential. None of the cell lines is tumorigenic. They should be interesting models for studying multistage carcinogenesis in human cells and transcriptional activation by TPA.
ISSN:0008-5472
1538-7445