In vitro and in vivo bioactivity of recombinant canine hepatocyte growth factor.
Hepatocyte growth factor (HGF) is crucial for the development and regeneration of the liver and offers a possible new therapeutic strategy for the treatment of canine liver disease. In this study, the in vitro and in vivo bioactivity of recombinant canine HGF (rcHGF) produced with a baculoviral expression system in insect cells was measured. In vitro rcHGF induced mitogenesis, motogenesis, and phosphorylated the HGF receptor c-MET and its downstream mediators PKB and ERK1/2 in two canine epithelial cell lines. After a partial hepatectomy (phx) in dogs, rcHGF increased phosphorylation of c-MET, PKB and ERK1/2. A moderate increase was seen with the cell cycle protein PCNA in rcHGF treated livers, but no HGF-induced increase in liver weight was seen 7 days after phx. Furthermore, rcHGF treated livers showed lower levels of the key mediator of apoptosis, caspase-3, at 7days after phx. It is concluded that rcHGF is a biologically active protein in vitro and in vivo and the baculoviral expression system supplies sufficient amounts of rcHGF for future clinical studies in dogs.
Arends
Brigitte
B
Faculty of Veterinary Medicine, Department of Clinical Sciences of Companion Animals, Utrecht University, Yalelaan 104, 3584 CM Utrecht, the Netherlands. B.Arends@vet.uu.nl
Spee
Bart
B
Hoffmann
Gaby
G
Jansen
Georgina E G
GE
Slump
Estel
E
Auriemma
Edoardo
E
Ijzer
Jooske
J
Hemrika
Wieger
W
Romijn
Roland A
RA
van der Heijden-Liefkens
Karin H A
KH
Sondermeijer
Paul J A
PJ
van den Ingh
Ted S G A M
TS
Penning
Louis C
LC
Rothuizen
Jan
J
D000818:Animals; D002460:Cell Line; D004285:Dogs; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006498:Hepatectomy; D017228:Hepatocyte Growth Factor / Q000235:genetics / Q000378:metabolism*; D022781:Hepatocytes / Q000187:drug effects*; D008099:Liver / Q000187:drug effects / Q000502:physiology; D008115:Liver Regeneration / Q000187:drug effects*; D009929:Organ Size; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D019859:Proto-Oncogene Proteins c-met / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins
D011994:Recombinant Proteins; D017228:Hepatocyte Growth Factor; D019859:Proto-Oncogene Proteins c-met; D051057:Proto-Oncogene Proteins c-akt; D048049:Extracellular Signal-Regulated MAP Kinases