Affinity with Raf is sufficient for Ras to efficiently induce rat mammary carcinomas.
The role of three major Ras downstream effector pathways in the induction of mammary cancer was studied using an in situ mammary ductal gene delivery model. Replication-defective retroviral vectors were used to infect endogenous rat mammary epithelial cells with three individual Ras effector loop mutants, each of which transduces its signal through a different Ras effector pathway (Raf, PI3K or RalGDS). Several groups have used Ras effector loop mutants in cultured cells, clearly characterizing the signaling specificity of each over a wide range of cell lines and conditions. Each of the three Ras effector loop mutations impairs Ras for neoplastic transformation of immortal cell lines in culture. In contrast, when evaluated in vivo by infecting endogenous rat mammary epithelial cells in situ with retroviral vectors, we find that codon 12 mutant activated V12-Ras and all three V12-Ras effector loop mutants individually induce mammary carcinomas. Most notably, a Ras effector loop mutant that lacks affinity with PI3K and RalGDS but retains affinity with Raf (E38-V12-Ras) is relatively similar in potency to V12-Ras for mammary carcinoma induction. Two other Ras effector loop mutants, each lacking affinity with Raf, one retaining affinity with PI3K (C40-V12-Ras), the other with RalGDS (G37-V12-Ras), resulted in much longer tumor latency than E38-V12-Ras and V12-Ras and a reduced carcinoma frequency. Tumor latencies for V12-Ras, E38-V12-Ras, C40-V12-Ras and G37-V12-Ras were 4, 4, 11 and 12 weeks, respectively. We conclude that the Ras-Raf pathway can function independently of the Ras-PI3K and Ras-RalGDS pathways for rapid induction of rat mammary carcinomas, while Ras-PI3K and Ras-RalGDS pathways may also individually induce mammary carcinomas following a long latency.
McFarlin
Daniel R
DR
McArdle Laboratory for Cancer Research and Department of Biostatistics and Medical Informatics, University of Wisconsin-Madison, 1400 University Avenue, Madison, WI 53706-1599, USA.
Lindstrom
Mary J
MJ
Gould
Michael N
MN
D016475:3T3 Cells; D000818:Animals; D001940:Breast / Q000378:metabolism / Q000473:pathology; D005822:Genetic Vectors; D015674:Mammary Neoplasms, Animal / Q000378:metabolism* / Q000473:pathology*; D051379:Mice; D009154:Mutation; D011485:Protein Binding; D019908:Proto-Oncogene Proteins c-raf / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D012190:Retroviridae; D013997:Time Factors; D018631:ras Proteins / Q000235:genetics / Q000378:metabolism*
D019908:Proto-Oncogene Proteins c-raf; D018631:ras Proteins