Suppression of pancreatic cancer by the dominant negative ras mutant, N116Y.
N116Y, H-ras mutant, possesses dominant negative activity to Ras function. The aim of this study is to assess whether N116Y can inhibit the proliferation of pancreatic cancer cell lines carrying K-ras mutations and cause reversion of the malignant phenotype. We transfected an expression vector of N116Y, pZIP-N116Y, into eight human pancreatic cancer cell lines with K-ras mutations (PCI 10, 19, 24, 35, 43, 55, 64, and 66) by using a lipofection procedure. The growth inhibition activity of N116Y was evaluated by the colony-forming efficiency in selection medium. In order to examine the effect of N116Y on the neoplastic phenotype, we established N116Y-expressing clones and analyzed their growth ability in soft agar and tumorigenicity in nude mice. The growth of the eight pancreatic cancer cell lines was strongly inhibited by the transfection of pZIP-N116Y. Moreover, the N116Y-expressing clones became less spread and lost their anchorage-independent growth ability. Furthermore, they were nontumorigenic in vivo. N116Y significantly inhibits the growth of pancreatic cancer cell lines and causes reversion of the malignant phenotypes. These results suggest that N116Y may be a candidate gene for use in the gene therapy of pancreatic cancer.
Shichinohe
T
T
Laboratory of Gene Regulation, Cancer Institute, Hokkaido University School of Medicine, Sapporo, Japan.
Senmaru
N
N
Furuuchi
K
K
Ogiso
Y
Y
Ishikura
H
H
Yoshiki
T
T
Takahashi
T
T
Kato
H
H
Kuzumaki
N
N
D000818:Animals; D000970:Antineoplastic Agents; D002277:Carcinoma / Q000235:genetics / Q000473:pathology*; D002455:Cell Division; D005799:Genes, Dominant; D006131:Growth Inhibitors; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000473:pathology*; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D006131:Growth Inhibitors; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)