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A genome-wide survey of RAS transformation targets

Nature Genetics volume 24, pages 144–152 (2000)Cite this article

Abstract

An important aspect of multi-step tumorigenesis is the mutational activation of genes of the RAS family, particularly in sporadic cancers of the pancreas, colon, lung and myeloid system1. RAS genes encode small GTP-binding proteins that affect gene expression in a global way by acting as major switches in signal transduction processes, coupling extracellular signals with transcription factors2,3,4. Oncogenic forms of RAS are locked in their active state and transduce signals essential for transformation, angiogenesis, invasion and metastasis via downstream pathways involving the RAF/MEK/ERK cascade of cytoplasmic kinases, the small GTP-binding proteins RAC and RHO, phosphatidylinositol 3-kinase and others5,6. We have used subtractive suppression hybridization (SSH), a PCR-based cDNA subtraction technique7, to contrast differential gene expression profiles in immortalized, non-tumorigenic rat embryo fibroblasts and in HRAS- transformed cells. Sequence and expression analysis of more than 1,200 subtracted cDNA fragments revealed transcriptional stimulation or repression of 104 ESTs, 45 novel sequences and 244 known genes in HRAS- transformed cells compared with normal cells. Furthermore, we identified common and distinct targets in cells transformed by mutant HRAS, KRAS and NRAS, as well as 61 putative target genes controlled by the RAF/MEK/ERK pathway in reverted cells treated with the MEK-specific inhibitor PD 98059.

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Figure 1: Characteristics of normal 208F fibroblasts (left) and HRAS-transformed FE-8 cells, untreated (middle) and incubated with the MEK-inhibitor PD 98059 (right).
Figure 2: Effects of the Ras/Raf/MEK signalling pathway and of different RAS isoforms on selected target gene transcription.
Figure 3: The 208F cell lines transformed with oncogenic HRAS, KRAS and NRAS exhibit similar neoplastic characteristics.

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Acknowledgements

We thank S. Petermann, J. Keil and K. Skultety for technical assistance; R. Brinckmann for sequencing; C. Schlüns for computing; F. Leenders and F. Theuring for help with phospho-image analysis; and M. Dietel for generous support. Our work was supported by Deutsche Krebshilfe (grant 10-332-Schä I to R.S.), Krebsliga des Kantons Zürich (grant to R.S.), Studienstiftung des Deutschen Volkes and Forschungsförderung der Charité (to J.Z.), and Berliner Krebsgesellschaft (grant to R.S.).

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  1. Johannes Zuber and Oleg I. Tchernitsa: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Molecular Tumour Pathology, Institute of Pathology, Charité, Humboldt-University, D-10117, Berlin, Germany

    Johannes Zuber, Oleg I. Tchernitsa, Anne-Chantal Schmitz, Martin Grips, Martin Hellriegel, Christine Sers & Reinhold Schäfer

  2. metaGen, Gesellschaft für Genomforschung mbH, D-14195, Berlin-Dahlem, Germany

    Bernd Hinzmann & André Rosenthal

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Correspondence to Reinhold Schäfer.

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Zuber, J., Tchernitsa, O., Hinzmann, B. et al. A genome-wide survey of RAS transformation targets. Nat Genet 24, 144–152 (2000). https://doi.org/10.1038/72799

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