Induction of putative tumor-suppressing genes in Rat-1 fibroblasts by oncogenic Raf-1 as evidenced by robot-assisted complex hybridization.
The growth factor receptor-dependent protein kinase Raf-1 is activated by GTP-bound Ras, thereby activating the mitogen-activated protein kinase pathway. To study the role of Raf in transformation we transduced Rat-1 cells with a tetracycline-regulatable retroviral vector encoding the constitutively active oncogenic C-terminal fragment of the human Raf-1 protein. Using subtractive hybridization of mRNAs from induced and noninduced cells and robot-assisted screening by complex hybridization, Raf-induced genes with various different characteristics of induction were investigated. Among the strongly induced genes were those involved in carcinogenesis such as metalloproteinases 3, 10 and 13, cathepsin L, ornithine decarboxylase, and putative tumor-suppressing genes such as monocyte chemoattracting protein 1, interferon-induced protein 10, a recently identified 2'-5' oligoadenylate synthetase-like protein, and plasminogen activator inhibitor type 2. Other components of the plasminogen activator system were not induced. Plasminogen activator inhibitor type 2 is a down-regulator of the proteolytic cascade consisting of various metalloproteinases, some of which are induced by a carboxy-terminal Raf mutant (RafCT). In conclusion, RafCT induces factors which act in a conflicting manner in respect of carcinogenesis, especially within the proteolytic system of the extracellular matrix.
Heinrich
J
J
Institute for Medical Virology, University of Zurich, Zuerich, Switzerland.
Bosse
M
M
Eickhoff
H
H
Nietfeld
W
W
Reinhardt
R
R
Lehrach
H
H
Moelling
K
K
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D056668:Cathepsin L; D002403:Cathepsins / Q000378:metabolism; D002460:Cell Line; D018932:Chemokine CCL2 / Q000378:metabolism; D054357:Chemokine CXCL10; D019743:Chemokines, CXC / Q000378:metabolism; D017364:Collagenases / Q000378:metabolism; D003546:Cysteine Endopeptidases; D010450:Endopeptidases; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D005419:Flavonoids / Q000494:pharmacology; D015723:Gene Library; D006801:Humans; D053506:Matrix Metalloproteinase 10; D053509:Matrix Metalloproteinase 13; D019278:Matrix Metalloproteinase 3 / Q000378:metabolism; D008666:Metalloendopeptidases / Q000378:metabolism; D008957:Models, Genetic; D009154:Mutation; D009693:Nucleic Acid Hybridization; D009955:Ornithine Decarboxylase / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D017396:Plasminogen Activator Inhibitor 2 / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D019908:Proto-Oncogene Proteins c-raf / Q000378:metabolism* / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D012190:Retroviridae / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013752:Tetracycline / Q000494:pharmacology; D013997:Time Factors; D015854:Up-Regulation
D018932:Chemokine CCL2; D054357:Chemokine CXCL10; D019743:Chemokines, CXC; D004791:Enzyme Inhibitors; D005419:Flavonoids; D017396:Plasminogen Activator Inhibitor 2; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D019908:Proto-Oncogene Proteins c-raf; C107753:proto-oncogene protein v-RAF1, pig; D002403:Cathepsins; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; C534281:CTSL protein, human; D056668:Cathepsin L; C534280:Ctsl protein, rat; D017364:Collagenases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13; D008666:Metalloendopeptidases; C507083:Mmp13 protein, rat; D019278:Matrix Metalloproteinase 3; D053506:Matrix Metalloproteinase 10; D009955:Ornithine Decarboxylase; D013752:Tetracycline; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one