Expression of fibroblast growth factor-1 (FGF-1), FGF-2 and FGF receptor-1 in a human salivary-gland adenocarcinoma cell line: evidence of growth.
Fibroblast growth factor-1 (FGF-1) and FGF-2 are heparin-binding polype ptides which express potent mitogenic properties in neoplastic cells. In the present study, we have examined the contribution of endogenous FGF-1 and FGF-2 to the autocrine growth of HSY human salivary-gland adenocarcinoma cells in vitro. Using specific monoclonal antibodies against FGF-1 and FGF-2, immunohistochemical analysis of HSY cells revealed strong expression of both FGF-1 and FGF-2 in the cytoplasm and nucleus. Consistent with these data, 2 molecular mass species of FGF-1 (16 and 18 kDa) and 3 FGF-2 (18, 24 and 27 kDa) were identified in HSY cells by Western-blot analysis. Scatchard analysis of FGF binding sites on HSY cells indicated the presence of 23,000 [125I]FGF-1 binding sites/cells with a dissociation constant (KD) of 178 pM and 13,000 [125I]FGF-2 binding sites/cell with a KD of 102 pM. In addition, HSY cells were shown to express the mRNA for FGF receptor-1 (FGFR-1) by reverse transcription-polymerase chain reaction (RT-PCR), confirming the existence of high-affinity FGF binding sites. The influence of endogenous FGF-1 and FGF-2 on HSY cell growth was evaluated by suppressing the expression and activity of FGF by using anti-sense oligonucleotides and neutralizing antibodies. The addition of 50 micron FGF-1-specific anti-sense oligonucleotides to HSY cells resulted in a 61% inhibition of cell growth, while 50 microM FGF-2-specific anti-sense oligonucleotides resulted in a 76% inhibition. These effects were dose-dependent and specific, since sense oligonucleotides were ineffective in inhibiting HSY cell growth at the same concentration. Furthermore, HSY cell growth was suppressed in the presence of anti-FGF-1 or anti-FGF-2 neutralizing antibody, resulting in a 58% inhibition at 8 micromilligrams/ml. Our observations suggest that FGF-1 and FGF-2 may act as autocrine regulators by interacting with FGF receptors on HSY cells.
Myoken
Y
Y
Department of Oral and Maxillofacial Surgery 1, Hiroshima University School of Dentistry, Japan.
Myoken
Y
Y
Okamoto
T
T
Kan
M
M
McKeehan
W L
WL
Sato
J D
JD
Takada
K
K
D000230:Adenocarcinoma / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D002455:Cell Division; D017931:DNA Primers / Q000737:chemistry; D016220:Fibroblast Growth Factor 1 / Q000502:physiology*; D016222:Fibroblast Growth Factor 2 / Q000502:physiology*; D015972:Gene Expression Regulation, Neoplastic; D005796:Genes; D006801:Humans; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger / Q000235:genetics; D012334:RNA, Neoplasm / Q000235:genetics; D017468:Receptors, Fibroblast Growth Factor / Q000502:physiology*; D012468:Salivary Gland Neoplasms / Q000235:genetics*
D017931:DNA Primers; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D017468:Receptors, Fibroblast Growth Factor; D016222:Fibroblast Growth Factor 2; D016220:Fibroblast Growth Factor 1