Hepatocyte growth factor transforms immortalized mouse liver epithelial cells.
Transforming activity of hepatocyte growth factor (HGF) was demonstrated utilizing immortalized but not fully transformed mouse hepatocytes (MLE-10). Rat HGF cDNA, expressed under the control of a cytomegalovirus promoter, was transfected together with the neomycin resistance gene (PSV2neo) into MLE-10 cells by the calcium phosphate method, and propagated G418-resistant colonies were harvested colony by colony. After checking for integration and expression of exogenous HGF, five cell lines (MLE-10-HGF-1-5) were established. Three cell lines transfected with the vector only (MLE-10-CMV-1-3) were also established in the same manner. All MLE-10-HGF cell lines grew much faster than the MLE-10-CMV and original MLE-10 cells in culture and produced large colonies in soft agar, which colony production was blocked by the addition of anti-HGF antibody to the agar. After addition of HGF, original and MLE-10-CMV lines produced colonies in soft agar. The high-HGF-production lines (MLE-10-HGF-4 and -5) also gave rise to tumors within 2 weeks when implanted into the nude mice subcutis. In contrast, all MLE-10-CMV and original MLE-10 cells were negative in these growth assays. A rough parallelism between the level of HGF expression and the growth rate in both soft agar and nude mice subcutis was evident among MLE-10-HGF cell lines. Those with higher HGF production tended to grow in a scattered fashion in culture. High-affinity HGF receptor, HGFR/met, was expressed in MLE-10 and all the derived cell lines. Since HGF and/or HGFR/met gene expression is seen in various tumors and the serum HGF level is elevated in patients with hepatic disease, the present results indicate a possible significance of HGF and its receptor system in carcinogenesis, most probably via autocrine and/or paracrine mechanisms.
Kanda
H
H
Department of Pathology, Cancer Institute, Tokyo, Japan.
Tajima
H
H
Lee
G H
GH
Nomura
K
K
Ohtake
K
K
Matsumoto
K
K
Nakamura
T
T
Kitagawa
T
T
D000818:Animals; D001483:Base Sequence; D002461:Cell Line, Transformed; D002465:Cell Movement / Q000187:drug effects; D002471:Cell Transformation, Neoplastic; D017228:Hepatocyte Growth Factor / Q000235:genetics / Q000502:physiology*; D008099:Liver / Q000166:cytology*; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D010957:Plasmids; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D019859:Proto-Oncogene Proteins c-met; D012333:RNA, Messenger / Q000032:analysis; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D017228:Hepatocyte Growth Factor; D019859:Proto-Oncogene Proteins c-met; D020794:Receptor Protein-Tyrosine Kinases