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Secretion of basic fibroblast growth factor (FGF-2) by WEHI-3B myelomonocytic leukemia cells
Augusto Pessina1, Giuseppina Gagliardi, Cristina Croera
1Institute of Microbiology, University of Milan, Via Pascal 36, 20133 Milan, Italy. pessinaa@mailserver.unimi.it
Abstract:
In order to investigate the role of Fibroblast Growth Factors in hematopoietic cells, we studied the expression of FGF-1, FGF-2, FGF-3, FGF-4, FGF-5 and FGF-6 mRNAs both in murine myelomonocytic leukemia WEHI-3B and in a murine stromal cell line SR-4987. Secretion of FGF-2 in the cell culture supernatant was also studied. Expression of mRNA encoding for the above-mentioned FGFs was analyzed by RT-PCR. The production of FGF-2 in the conditioned media of WEHI-3B and SR-4987 cell cultures was evaluated by techniques of affinity chromatography, chromatofocusing and immunoblotting. The biological activity of FGF-2 was checked on SR-4987 cells by a agar clonogenic assay. In both cell lines mRNA was found encoding for FGF-1, FGF-2 and FGF-6 and WEHI-3B cells express also mRNA for FGF-3 (int-2) and FGF-4 (K-FGF/hst). Furthermore, supernatant from WEHI-3B cells was found to stimulate dramatically the agar clonogenicity of SR-4987 cells which have a very poor basal capacity for growth in agar. The clonogenic activity of WEHI-3B conditioned medium is due to FGF-2 secreted into cell culture supernatant whereas SR-4987 cells, although express FGF-2 mRNA, do not seem able to secrete this factor. The expression in myeloid leukemia cells of oncogene-related factors such as FGF-3, FGF-4 and FGF-6 together with the secretion of FGF-2 able to support a positive regulation of bone marrow stromal cells function suggest that FGFs may have an important role in sustaining the leukemogenic process and related disorders.
Insights
Fibroblast Growth Factors (FGFs) are expressed in leukemia and stromal cells. Secreted FGF-2 from leukemia cells stimulates stromal cell growth, suggesting FGFs support leukemogenesis.
Area of Science:
- Hematology
- Molecular Biology
- Cancer Research
Background:
- Fibroblast Growth Factors (FGFs) are crucial signaling molecules.
- Their role in hematopoietic and stromal cells, particularly in leukemia, requires further investigation.
Purpose of the Study:
- To investigate the expression and function of various FGFs (FGF-1 to FGF-6) in murine leukemia (WEHI-3B) and stromal (SR-4987) cell lines.
- To determine the role of secreted FGF-2 in supporting stromal cell function and its potential contribution to leukemogenesis.
Main Methods:
- Reverse Transcription Polymerase Chain Reaction (RT-PCR) for mRNA expression analysis.
- Affinity chromatography, chromatofocusing, and immunoblotting for FGF-2 detection in cell culture supernatants.
- Agar clonogenic assay to assess the biological activity of FGF-2 on stromal cells.
Main Results:
- Both WEHI-3B and SR-4987 cells express mRNA for FGF-1, FGF-2, and FGF-6.
- WEHI-3B cells also express mRNA for FGF-3 and FGF-4.
- Supernatant from WEHI-3B cells significantly enhanced the agar clonogenicity of SR-4987 cells, mediated by secreted FGF-2.
- SR-4987 cells express FGF-2 mRNA but do not appear to secrete it.
Conclusions:
- FGFs, including oncogene-related factors like FGF-3, FGF-4, and FGF-6, are expressed in myeloid leukemia cells.
- Secreted FGF-2 from leukemia cells positively regulates bone marrow stromal cell function.
- FGFs likely play a significant role in sustaining the leukemogenic process and related hematological disorders.