Growth and Phenotoypic Characteristics of Human Nevus Cells in Culture

Mancianti, Maria Laura; Herlyn, Meenhard; Weil, Deborah; Jambrosic, J.; Rodeck, Ulrich; Becker, Dorothea; Diamond, Leila; Clark, Wallace H.; Koprowski, Hilary

doi:10.1111/1523-1747.ep12462099

Cited by 51 publications

(30 citation statements)

References 22 publications

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“…Cell growth was determined after 3 or 6 days by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay as described (Mosmann 1983) with minor modi®cations. Soft-agar assays for anchorage-independent growth were performed as described (Mancianti et al 1988). Brie¯y, a 0.5% agar layer in W489 medium containing 2% FCS was prepared in 35-mm wells.…”

Section: Melanocyte/melanoma Growth Assaysmentioning

confidence: 99%

Melanoma-associated expression of vascular endothelial growth factor and its receptors FLT-1 and KDR

Graeven¹,

Fiedler

Karpinski³

et al. 1999

Journal of Cancer Research and Clinical Oncology

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The expression patterns of vascular endothelial growth factor (VEGF) and its two receptors, flt-1 and KDR, were assessed in normal human melanocytes, transformed melanocytes expressing the simian virus 40 Tgene (SV40T), and melanoma cells derived from primary and metastatic lesions. Constitutive expression of VEGF, flt-1, and KDR mRNA and proteins was observed in the majority of primary and metastatic melanoma cell lines, and in SV40T-transformed melanocytes. VEGF expression in melanoma cell lines was further enhanced by exogenous growth factors including insulin and fetal calf serum. By contrast, neonatal melanocytes did not express VEGF or VEGF receptors and VEGF expression could not be induced by exogenous growth factors. Exogenous VEGF had no significant effects on melanoma cell proliferation or on production of a transcriptional target for VEGF, urokinase-type plasminogen activator. Down-regulation of VEGF expression in the metastatic melanoma cell line WM164 through transfection of a VEGF antisense construct similarly did not affect proliferation of the transfected cells in the presence or absence of exogenous VEGF. In summary, coexpression of VEGF and its receptors is a tumor-associated phenomenon in melanoma development. However VEGF production does not support autocrine proliferation of the melanoma cell lines tested.

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Section: Melanocyte/melanoma Growth Assaysmentioning

confidence: 99%

Melanoma-associated expression of vascular endothelial growth factor and its receptors FLT-1 and KDR

Graeven¹,

Fiedler

Karpinski³

et al. 1999

Journal of Cancer Research and Clinical Oncology

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“…The procedure for isolating melanocytic cells from normal skin, benign melanocytic lesions, and primary melanomas consists of separating the epidermal portion of the skin from the dermal portion using trypsin, followed by further disaggregation of each layer of cells with the enzymes, dispase, collagenase, and hyaluronidase [9]. Improved selective media to counter contamination by fibroblasts and keratinocytes can yield pure melanocytic cultures.…”

Section: Culture Of Melanocytic Cellsmentioning

confidence: 99%

Human melanoma: Development and progression

1990

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Clinical and histopathological evidence suggests that melanoma develops in a sequence of steps, progressing from benign proliferative lesions, to primary melanomas that do not show evidence for metastasis, to invasive primary lesions, and to metastases. This review focuses on the experimental studies examining the phenotypic characteristics of cultured primary melanoma cells as they relate to cells from non-malignant nevi and metastases. Genetic, biologic, and immunologic criteria have been established to distinguish melanocytes from different steps of tumor development. These include non-random chromosomal abnormalities, expression of melanocyte- and melanoma-specific antigens, requirements for exogenous growth factors, production of endogenous growth factors, and expression of receptors for growth factors. The transformation of melanocytes and nevus cells with viral oncogenes has facilitated studies on the malignant phenotype. Variants have been developed through successive selections from primary melanoma cell populations that have one or several characteristics of metastatic cells. The study of melanocytes isolated from various stages of tumor development and the generation of cell variants with specific properties should enable a long-term search for the molecular mechanisms of melanoma development and progression.

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“…Rodeck et al [40] and Mancianti et al [41] postulated the loss of sensitivity to growth inhibitory factors as another contributor of growth factor independence in metastatic tumor cells. They found a stage dependent subsequent loss of exogenous growth factor requirements in human melanoma cell lines.…”

Section: Discussionmentioning

confidence: 99%

Aberrations of growth factor control in metastatic follicular thyroid cancerin vitro

Hoelting¹,

Zielke²,

Siperstein³

et al. 1994

Clin Exp Metast

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The aggressiveness of follicular thyroid cancer (FTC) varies widely, and metastasis is the primary cause of death. Uncontrolled proliferation of cancer cells may be associated with loss of growth factor control. We investigated the effects of stimulating (epidermal growth factor [EGF]; thyreotropin [TSH] in low concentrations) and inhibiting growth factors (transforming growth factor beta 1 [TGF beta 1]; TSH in high concentrations) on invasion and growth of FTC cell lines from the thyroid tumor (FTC133) and from the lymph node (FTC236) and lung (FTC238) metastases of the same patient. Invasion-penetration through an 8 microns pore membrane, covered by Matrigel (basement membrane)-and growth were measured using the MTT-method. EGF (10 ng/ml) and TSH in low concentrations (1 mU/ml) stimulated invasion and growth of all FTC cell lines, but the amplitude of stimulation differed significantly. The parental cell line FTC133 was considerably more responsive to growth factor stimulation than the metastatic clones. Invasion of FTC133 was enhanced by 42% (EGF; p < 0.02) and 21% (TSH; p < 0.01), invasion of FTC236 by 8% (EGF; p < 0.02) and 8% (TSH; p < 0.01), and invasion of FTC238 by 9% (EGF; p < 0.02) and 8% (TSH; p < 0.01). Conversely, invasion and growth of FTC133 were significantly more inhibited by TGF beta 1 (10 ng/ml) and supraphysiologic concentrations of TSH (100 mU/ml) than the cell lines from the lymph node and lung metastases. At day 7, invasion of FTC133 was inhibited by 32% (TGF beta 1; p < 0.02) and 21% (TSH; p < 0.01), invasion of FTC236 by 18% (TGF beta 1; p < 0.02) and 11% (TSH; p < 0.01), and invasion of FTC238 by 16% (TGF beta 1; p < 0.02) and 12% (TSH; p < 0.01). Moreover, we analyzed growth factor independence in minimally supplemented or unsupplemented medium. Growth, but no invasion was evident when cells were cultured completely unsupplemented over 7 days. These results suggest that metastatic FTCs may have developed by escaping from the normal control of TSH and other growth factors.

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Growth and Phenotoypic Characteristics of Human Nevus Cells in Culture

Cited by 51 publications

References 22 publications

Melanoma-associated expression of vascular endothelial growth factor and its receptors FLT-1 and KDR

Melanoma-associated expression of vascular endothelial growth factor and its receptors FLT-1 and KDR

Human melanoma: Development and progression

Aberrations of growth factor control in metastatic follicular thyroid cancerin vitro

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