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Elevated level of p60c-src in virus-transformed murine megakaryocytic cell lines
S Fischer1, F Wendling, I Boulet
1Unité 15 INSERM, CHU Cochin, Paris, France.
Abstract:
Megakaryocytic cell lines derived from mouse bone marrow cells transformed by the Myeloproliferative Leukemia Virus (MPLV) contain elevated levels of p60c-src. Northern blot analysis revealed the presence of a 4 kb normal sized c-src transcript only in MPLV-transformed megakaryocytic cell lines containing a high percentage of acetylcholinesterase positive cells (AChE+ greater than 10%), but not in MPLV-transformed erythroblastic or myeloblastic cell lines. The p60c-src protein was identified in lysates from in vivo labelled cells and in in vitro labelled membrane extracts by immunoblotting analysis and by immunoprecipitations with specific anti-src antibodies. In dimethylsulfoxide (DMSO) treated cells, the number of AChE+ cells increased together with p60c-src kinase activity indicating a possible correlation between p60c-src expression/activity and megakaryocytic differentiation.
Insights
Myeloproliferative Leukemia Virus (MPLV) transformed megakaryocytic cells show increased p60c-src levels. This suggests a correlation between p60c-src activity and megakaryocytic differentiation.
Area of Science:
- Molecular Biology
- Cell Biology
- Hematopoiesis
Background:
- The Myeloproliferative Leukemia Virus (MPLV) can transform hematopoietic cells.
- The c-src proto-oncogene encodes a non-receptor tyrosine kinase, p60c-src, implicated in cell growth and differentiation.
Purpose of the Study:
- To investigate the role of p60c-src in MPLV-induced megakaryocytic differentiation.
- To determine if p60c-src expression or activity correlates with megakaryocytic cell maturation.
Main Methods:
- Northern blot analysis to detect c-src transcripts.
- Immunoblotting and immunoprecipitation to identify and quantify p60c-src protein.
- Acetylcholinesterase (AChE) staining to assess megakaryocytic differentiation.
- Dimethylsulfoxide (DMSO) treatment to induce differentiation.
Main Results:
- MPLV-transformed megakaryocytic cell lines exhibited elevated p60c-src levels compared to other cell types.
- A normal-sized c-src transcript was detected specifically in megakaryocytic lines with high AChE activity.
- p60c-src kinase activity increased with DMSO-induced megakaryocytic differentiation (AChE+ cells).
Conclusions:
- p60c-src expression and kinase activity are associated with megakaryocytic differentiation in MPLV-transformed cells.
- Elevated p60c-src may play a role in the maturation of megakaryocytes.
- Further research is warranted to elucidate the precise mechanism of p60c-src in megakaryopoiesis.