Tracheal Smooth Muscle Cells Stimulated by Stem Cell Factor-c-Kit Coordinate the Production of Transforming Growth Factor-β1 and Fibroblast Growth Factor-2 Mediated by Chemokine (C-C Motif) Ligand 3.
The aim of this study was to evaluate the mechanism involved in the stem cell factor (SCF)-induced production of fibroblast growth factor-2 (FGF-2), transforming growth factor-β1 (TGF-β1), and chemokine (C-C motif) ligand 3 (CCL3) in tracheal smooth muscle cells (tSMCs) and the signaling pathway involved in the process. tSMC primary cultures were stimulated with SCF and evaluated at 24 h. Cells treated with specific antibodies did not show any immunolabeling for cytokeratin or fibroblast activation protein, but were positive for α-smooth muscle actin, indicating the purity of the primary cell line. Western blot analysis showed constitutive phosphorylation of c-Kit, as well as increased total protein and phosphorylated c-Kit levels in tSMCs after SCF stimulation. Flow cytometry analysis also showed an increase in cell-surface c-Kit expression in the presence of SCF. SCF induced TGF-β mRNA expression in tSMCs, as well as the production of TGF-β1, CCL3, and FGF-2. Pretreatment with anti-CCL3 antibody blocked TGF-β1 expression and partially inhibited FGF-2 production. On the other hand, anti-c-Kit antibody blocked TGF-β1 expression and FGF-2 production. Thus, TGF-β1 and FGF-2 production were mediated by CCL3 production through c-Kit. Pretreatment with mitogen-activated protein kinase kinase 1, p38, and Jun N-terminal kinase inhibitors showed that the effects mediated by SCF were involved with the modulation of mitogen-activated protein kinase (MAPK) pathways. Development of inhibitors targeting CCL3 through MAPK activation could thus be an attractive strategy to inhibit tSMC activation during asthma.
1 Programa de Pós-graduação Multicêntrico em Ciências Fisiológicas-SBFis, Department of Basic Sciences, School of Dentistry of Araçatuba, Univ. Estadual Paulista-UNESP , Araçatuba, Brazil .
Luis Cezar Farias de
LC
Oliveira
1 Programa de Pós-graduação Multicêntrico em Ciências Fisiológicas-SBFis, Department of Basic Sciences, School of Dentistry of Araçatuba, Univ. Estadual Paulista-UNESP , Araçatuba, Brazil .
Taís Marolato
TM
Danilucci
1 Programa de Pós-graduação Multicêntrico em Ciências Fisiológicas-SBFis, Department of Basic Sciences, School of Dentistry of Araçatuba, Univ. Estadual Paulista-UNESP , Araçatuba, Brazil .
Antonio Hernandes
AH
Chaves-Neto
2 Department of Biological Sciences, School of Dentistry of Bauru, São Paulo University-USP , Bauru, Brazil .
Ana Paula
AP
Campanelli
3 Laboratory of Animal Virology and Cell Culture, School of Medicine Veterinary of Araçatuba, Univ. Estadual Paulista-UNESP , Araçatuba, Brazil .
Tereza Cristina Cardoso da
TC
Silva
1 Programa de Pós-graduação Multicêntrico em Ciências Fisiológicas-SBFis, Department of Basic Sciences, School of Dentistry of Araçatuba, Univ. Estadual Paulista-UNESP , Araçatuba, Brazil .
Sandra Helena Penha
SH
Oliveira
D000818:Animals; D054405:Chemokine CCL3 / Q000235:genetics / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis* / Q000235:genetics; D015870:Gene Expression; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D008297:Male; D051379:Mice; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism*; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D019089:Stem Cell Factor / Q000378:metabolism* / Q000494:pharmacology; D014132:Trachea / Q000166:cytology*; D053773:Transforming Growth Factor beta1 / Q000096:biosynthesis* / Q000235:genetics; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
D054405:Chemokine CCL3; D012333:RNA, Messenger; D019089:Stem Cell Factor; D053773:Transforming Growth Factor beta1; D016222:Fibroblast Growth Factor 2; D019009:Proto-Oncogene Proteins c-kit; D048031:JNK Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases