[The role of human fibroblast growth factor receptor 1-IIIb isoform in proliferation of pancreatic ductal cells and its effects to mitogen-activated protein kinase].
OBJECTIVE
To study the role of IIIb isoform of human fibroblast growth factor receptor 1 (FGFR1-IIIb) in proliferation of pancreatic ductal cells and its effects on mitogen-activated protein kinase (MAPK).


METHODS
Human pancreatic ductal cells of the line TAKA-1 were cultured. The plasmid of human full-length FGFR1-IIIb isoform, pSVK4/FGFR1-IIIb, was stable transfected into the cultured TAKA-1 pancreatic ductal cells facilitated by lipofectamine. Un-transfected TAKA-1 cells and TAKA-1 ductal cells transfected with blank plasmid were used as controls. The expression, distribution and character of protein of FGFR1-IIIb in the TAKA-1 cells were estimated by Western blotting, Northern blotting, immunofluorescence assay, and glycosylation assay. The function and mechanism of FGFR1-IIIb in the transfected pancreatic ductal cells stimulated by FGF were examined by MTT assay and MAPK assay. Tunicamycin, an inhibitor of N-terminal glycoprotein synthesis, was added into the culture fluid of the FGFR1-IIIb transfected TAKA-1 cells to observe the changes of the FGFR1 bands.


RESULTS
FGFR1-IIIb, a glycosylated receptor at various levels at 120 kDa and between 130 - 150 kDa, was localized at moderate levels at the cell membrane and cytoplasm and at higher level in the perinuclear region of the cytoplasm of the pSVK4/FGFR1-IIIb-transfected cells. FGF-1, -2, and -4 significantly increased the growth of FGFR1-IIIb-transfected TAKA-1 cells, and at the same time induced the p44/p42 MAPK phosphorylation.


CONCLUSIONS
Human FGFR1-IIIb receptor is a functional receptor in pancreatic ductal cells. FGF-1, -2, and -4 can increase the growth of FGFR1-IIIb-transfected pancreatic ductal cells, and the mechanism is that they can induce the p44/p42 MAPK phosphorylation.
Liu
Zhan-bing
ZB
Department of General Surgery, Peking University First Hospital, Peking University, Beijing 100034, China. zhanbing03@yahoo.com.cn
Yang
Yin-mo
YM
Qiao
Qi-lu
QL
Wan
Yuan-lian
YL
Kornmann
Marko
M
Huang
Yan-ting
YT
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D054638:Dual Specificity Phosphatase 1 / Q000378:metabolism*; D016220:Fibroblast Growth Factor 1 / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D006168:Guinea Pigs; D006801:Humans; D010183:Pancreatic Ducts / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D010957:Plasmids; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D051496:Receptor, Fibroblast Growth Factor, Type 1 / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D020033:Protein Isoforms; D016222:Fibroblast Growth Factor 2; D016220:Fibroblast Growth Factor 1; D051496:Receptor, Fibroblast Growth Factor, Type 1; D054638:Dual Specificity Phosphatase 1