Release of fibroblast growth factor-1 by human squamous cell carcinoma correlates with autocrine cell growth.
A squamous cell carcinoma cell line Nakata proliferated in serum-free culture and was not responsive to exogenous fibroblast growth factor-1 (FGF-1). Immunostaining revealed that Nakata cells expressed FGF-1 in their cytoplasms and nuclei. Two molecular mass species of FGF-1 (16 and 18 kDa) were identified in cell extracts by Western blot. These cells also expressed high-affinity FGF-1 binding sites (Kd = 360 pM, 28,000 sites/cell). The results of cross-linking with [125I]FGF-1 demonstrated the presence of two bands with molecular masses of 160 and 140 kDa. The addition of FGF-1 specific antisense oligonucleotides at 25 microM to Nakata cells resulted in an 82% inhibition in cell growth and suppressed FGF-1 expression. This effect was dose-dependent and specific, because sense oligonucleotides were ineffective in inhibiting cell growth. In addition, Nakata cell growth was suppressed by an anti-FGF-1 neutralizing antibody, which resulted in a 52% inhibition at 8 micrograms/ml. These results demonstrate that Nakata cells produce FGF-1, and indicate that this growth factor acts in an autocrine manner by interacting with FGF-1 binding sites on Nakata cells.
Myoken
Y
Y
Department of Oral and Maxillofacial Surgery, Hiroshima University School of Dentistry, Japan.
Myoken
Y
Y
Okamoto
T
T
Kan
M
M
Sato
J D
JD
Takada
K
K
D001483:Base Sequence; D015153:Blotting, Western; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D002455:Cell Division / Q000502:physiology; D016895:Culture Media, Serum-Free; D017931:DNA Primers / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D004273:DNA, Neoplasm / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D016220:Fibroblast Growth Factor 1 / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008969:Molecular Sequence Data; D009062:Mouth Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D016376:Oligonucleotides, Antisense / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D014407:Tumor Cells, Cultured
D016895:Culture Media, Serum-Free; D017931:DNA Primers; D004273:DNA, Neoplasm; D016376:Oligonucleotides, Antisense; D016220:Fibroblast Growth Factor 1