Secretion of basic fibroblast growth factor (FGF-2) by WEHI-3B myelomonocytic leukemia cells.
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.
Pessina
Augusto
A
Institute of Microbiology, University of Milan, Via Pascal 36, 20133 Milan, Italy. pessinaa@mailserver.unimi.it
Gagliardi
Giuseppina
G
Croera
Cristina
C
Foti
Paola
P
Dassi
Cristina
C
Brambilla
Paolo
P
Neri
Maria Grazia
MG
D000818:Animals; D002478:Cells, Cultured; D002845:Chromatography; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D016220:Fibroblast Growth Factor 1 / Q000096:biosynthesis; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis / Q000378:metabolism*; D051518:Fibroblast Growth Factor 3; D051519:Fibroblast Growth Factor 4; D051522:Fibroblast Growth Factor 6; D005346:Fibroblast Growth Factors / Q000096:biosynthesis; D015151:Immunoblotting; D007938:Leukemia / Q000378:metabolism*; D051379:Mice; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D017077:Culture Media, Conditioned; C489197:Fgf3 protein, mouse; C496446:Fgf4 protein, mouse; C486115:Fgf6 protein, mouse; D051518:Fibroblast Growth Factor 3; D051519:Fibroblast Growth Factor 4; D051522:Fibroblast Growth Factor 6; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D016222:Fibroblast Growth Factor 2; D016220:Fibroblast Growth Factor 1; D005346:Fibroblast Growth Factors