Mouse liver
tumors frequently harbor activating ras gene mutations. Downstream effector molecules of p21Ras include
Raf-1 kinase which mediates external signals via
kinase signaling pathways to nuclear
transcription factors including c-Fos and c-Jun. Mouse liver
tumors with differing ras-mutational status were analyzed for alterations in Ras/Raf-1 signal transduction.
Tumors were characterized with respect to the presence of base substitutions in the 3 known hot-spot positions at
codons 12, 13, and 61 of Ha-ras, Ki-ras, and N-ras. Ha-ras
codon 61 or Ki-ras
codon 13 mutations, but no N-ras mutations, were detected in 23 out of 33
tumors analyzed, while no ras-mutations were found in 10 of the
tumors. There was no significant difference in the expression of p21RaS
proteins between ras-mutated
tumors and
tumors without detectable ras mutations. To allow for determination of
Raf-1 kinase activity in
tumors, a sensitive and specific assay was developed for measurements with tissue homogenates.
Raf-1 kinase activity was increased about four-fold in liver
tumors as compared with normal liver tissue. No significant differences in
kinase activity, however, were evident between ras-mutated and ras-wild-type
tumors. The same was true with respect to the levels of c-fos and c-jun mRNAs. Moreover, there were no significant differences in cell division (5-bromo-2'-deoxyuridine-labeling indices) of hepatocytes from ras-mutated and ras-wild-type
tumors. The similar degree of constitutive activation of the Ras/Raf-1 signaling pathway in liver
tumors, with and without detectable ras mutations, suggests that other molecules within the signaling pathway may substitute for ras-mutations during oncogenic conversion of ras-wild-type hepatocytes.