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https://hdl.handle.net/2440/9108
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dc.contributor.author | Caruana, G. | - |
dc.contributor.author | Cambarer, A. | - |
dc.contributor.author | Gonda, T. | - |
dc.contributor.author | Ashman, L. | - |
dc.date.issued | 1998 | - |
dc.identifier.citation | Oncogene, 1998; 16(2):179-190 | - |
dc.identifier.issn | 0950-9232 | - |
dc.identifier.issn | 1476-5594 | - |
dc.identifier.uri | http://hdl.handle.net/2440/9108 | - |
dc.description.abstract | Ectopic expression of the normal murine receptor tyrosine kinase, c-Kit, in NIH3T3 cells induced many phenotypic changes characteristic of transformation including anchorage-independent growth, focus formation and tumorigenicity in nude mice. Although transformation was largely dependent on the presence of recombinant murine Steel Factor (SLF), the ligand to the c-Kit receptor, anchorage independent growth did occur at a low frequency in the absence of added factor, and this could not be inhibited by neutralising antibodies or by SLF anti-sense mRNA. Clones from factor-independent colonies in semi-solid agar displayed a narrow range of c-Kit surface protein levels (4.3-6.4 x 10(4) receptors/cell) which was relatively high compared with the pool from which they were derived. Analysis of a larger series of random clones derived from adherent cultures expressing different levels of c-Kit demonstrated a positive correlation between SLF-dependent, anchorage-independent growth and c-Kit protein and mRNA expression levels (respectively, Rs = 0.58, P < 0.01; and Rs = 0.53, P < 0.01) with consistent colony formation observed with clones having > 2.5 x 10(4) receptors/cell. Interestingly, two of the three clones expressing the highest levels of c-Kit protein and mRNA produced few or no colonies in the presence or absence of SLF. Sequential overexpression of human c-KIT in NIH3T3 cells using a dihydrofolate reductase (DHFR)-encoding vector and gene co-amplification through methotrexate selection, which resulted in pools expressing up to 1.5 x 10(5) receptors/cell, confirmed that high receptor densities resulted in a decrease in colony numbers. Thus, analysis of clonal and selected populations has indicated that an optimal level of c-Kit is required for transformation of NIH3T3 cells in the presence of SLF, and that some ligand-independent transformation occurs. | - |
dc.language.iso | en | - |
dc.publisher | STOCKTON PRESS | - |
dc.source.uri | http://dx.doi.org/10.1038/sj.onc.1201494 | - |
dc.subject | 3T3 Cells | - |
dc.subject | Animals | - |
dc.subject | Mice | - |
dc.subject | Cell Transformation, Neoplastic | - |
dc.subject | Methotrexate | - |
dc.subject | Ligands | - |
dc.subject | Cell Division | - |
dc.subject | Proto-Oncogene Proteins c-kit | - |
dc.title | Transformation of NIH3T3 fibroblasts by the c-kit receptor tyrosine kinase; effect of receptor density and ligand-requirement | - |
dc.type | Journal article | - |
dc.identifier.doi | 10.1038/sj.onc.1201494 | - |
pubs.publication-status | Published | - |
dc.identifier.orcid | Gonda, T. [0000-0002-8792-3021] | - |
Appears in Collections: | Aurora harvest 4 Medicine publications |
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