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Expression of fibroblast growth factor‐1 (FGF‐1), FGF‐2 and FGF receptor‐1 in a human salivary‐gland adenocarcinoma cell line: Evidence of autocrine growth
Author(s) -
Myoken Yoshiko,
Myoken Yoshinari,
Okamoto Tetsuji,
Kan Mikio,
McKeehan Wallace L.,
Sato J. Denry,
Takada Kazuaki
Publication year - 1996
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/(sici)1097-0215(19960301)65:5<650::aid-ijc15>3.0.co;2-b
Subject(s) - fibroblast growth factor , autocrine signalling , fibroblast growth factor receptor 1 , fibroblast growth factor receptor 4 , fibroblast growth factor receptor , fibroblast growth factor receptor 3 , cancer research , endocrinology , medicine , fibroblast growth factor 23 , fibroblast growth factor receptor 2 , biology , receptor , parathyroid hormone , calcium
Fibroblast growth factor‐1 (FGF‐1) and FGF‐2 are heparin‐binding polypeptides 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 [ 125 I] FGF‐1 binding sites/cells with a dissociation constant (K D ) of 178 pM and 13,000 [ 125 I]FGF‐2 binding sites/cell with a K D of 102 pM. In addition, HSY cells were shown to express the mRNA for FGF receptor‐I (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 μM FGF‐1‐specific anti‐sense oligonucleotides to HSY cells resulted in a 61% inhibition of cell growth, while 50 μM 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 μmg/ml. Our observations suggest that FGF‐1 and FGF‐2 may act as autocrine regulators by interacting with FGF receptors on HSY cells. © 1996 Wiley‐Liss, Inc.