Src/ERK but not phospholipase D is involved in keratinocyte growth factor-stimulated secretion of matrix metalloprotease-9 and urokinase-type plasminogen activator in SNU-16 human stomach cancer cell.
OBJECTIVE
We investigated the signaling pathway for keratinocyte growth factor (KGF)-induced invasion using human stomach cancer cell line, SNU-16.


METHODS
Alterations in the activities of Src, extracellular signal-regulated kinase (ERK), and phospholipase D (PLD) were measured using [gamma-(32)P] ATP for autophosphorylation of Src, phospho-specific ERK antibody, and [9,10-(3)H] myristic acid, respectively, while herbimycin A, PD98059 and butan-1-ol were used to inhibit their activities. Matrix metalloproteases (MMPs) and urokinase-type plasminogen activator (uPA) were quantified with zymography and Matrigel-coated Transwell was employed to estimate the invasiveness of SNU-16 cells.


RESULTS
Src, ERK, and PLD were activated in response to KGF treatment, and inhibition of these enzymes - by their specific inhibitors - decreased KGF-induced invasion in a dose-dependent manner. However, only inhibition of Src and ERK could block KGF-stimulated secretion of uPA and MMP-9.


CONCLUSIONS
Src, ERK, and PLD are suggested as mediators of KGF-induced invasion in SNU-16. uPA and MMP-9 are considered as downstream targets of Src and ERK whereas PLD is thought to utilize different pathways.
Shin
Eun-Young
EY
Department of Biochemistry, College of Medicine, Medical Research Institute, Chungbuk National University, San 48, Gaesin-dong, Heungduk-ku, Cheongju, 361-763, Korea.
Ma
Eun-Kyung
EK
Kim
Chi-Kyung
CK
Kwak
Sahng-June
SJ
Kim
Eung-Gook
EG
D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D003094:Collagen / Q000737:chemistry; D004338:Drug Combinations; D004791:Enzyme Inhibitors / Q000494:pharmacology; D051523:Fibroblast Growth Factor 7; D005346:Fibroblast Growth Factors / Q000494:pharmacology*; D005419:Flavonoids / Q000494:pharmacology; D006801:Humans; D007797:Laminin / Q000737:chemistry; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D010739:Phospholipase D / Q000378:metabolism*; D010766:Phosphorylation; D011509:Proteoglycans / Q000737:chemistry; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000378:metabolism*; D051497:Receptor, Fibroblast Growth Factor, Type 2; D017468:Receptors, Fibroblast Growth Factor / Q000378:metabolism; D015398:Signal Transduction; D013274:Stomach Neoplasms / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D014407:Tumor Cells, Cultured / Q000187:drug effects; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
D004338:Drug Combinations; D004791:Enzyme Inhibitors; C496453:FGF7 protein, human; D005419:Flavonoids; D007797:Laminin; D011509:Proteoglycans; D017468:Receptors, Fibroblast Growth Factor; C059485:matrigel; D051523:Fibroblast Growth Factor 7; D005346:Fibroblast Growth Factors; D003094:Collagen; D051497:Receptor, Fibroblast Growth Factor, Type 2; C065104:keratinocyte growth factor receptor; D016392:Proto-Oncogene Proteins pp60(c-src); D017871:Calcium-Calmodulin-Dependent Protein Kinases; D020928:Mitogen-Activated Protein Kinases; D010739:Phospholipase D; D014568:Urokinase-Type Plasminogen Activator; D020780:Matrix Metalloproteinase 9; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one