Measurement of c-myc protein content and cell cycle kinetics of normal and spontaneously transformed murine mastocytes by bivariate flow cytometry
| Autor: | Elio Geido, A. Di Vinci, B. Marsano, W. Giaretti, M Minks, Silvia Bruno |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 1990 |
| Předmět: |
medicine.drug_class
Monoclonal antibody Flow cytometry Proto-Oncogene Proteins c-myc chemistry.chemical_compound Mice medicine Animals Mast Cells Cell Line Transformed Cell Nucleus medicine.diagnostic_test biology Cell Cycle Antibodies Monoclonal Cell Biology General Medicine DNA Cell cycle Flow Cytometry Molecular biology Kinetics chemistry Bromodeoxyuridine Cell culture Mice Inbred DBA Monoclonal Cell Cycle Kinetics biology.protein Female Antibody Propidium |
| Popis: | Progressive in vitro culturing of interleukin-3 (IL-3) dependent normal murine mastocytes (PB-3) resulted in a variant cell line (PB-1) able to grow without exogenous IL-3 and which was tumorogenic in syngenic mice. Bivariate flow cytometry was used to evaluate the c-myc protein and DNA content of PB-3 and PB-1 cells. The c-myc protein was detected by specific monoclonal antibodies. Kinetic characteristics of PB-3 and PB-1 cell lines, namely, the duration of the G1, S and G2 + M cell cycle phases were also evaluated using the bromodeoxyuridine (BrdU) pulse-chase method and BrdU/DNA flow cytometry. Levels of c-myc protein in PB-1 cells were about two-fold higher than those of PB-3 cells in all cell cycle phases. Mean duration of the cell cycle (Tc) was 15.3 h for PB-3 cells and 12.4 h for PB-1 cells. Shortening in Tc for the transformed cells was due to a decrease of nearly 30% in mean duration of the G1 phase (from 8 h to 5.7 h). No significant differences were found in the duration of the S and G2 + M phases. These results indicate that acquired IL-3 independency in vitro and tumorogenicity of PB-1 cells were accompanied by a doubling of c-myc protein level and by a parallel shortening, or bypass, of the regulatory events within the G1 phase of the cell cycle. |
| Databáze: | OpenAIRE |
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