Abstract
RAS is a small GTP binding protein mutated in approximately 30% human cancer. Despite its important role in the initiation and progression of human cancer, the underlying mechanism of RAS-induced human epithelial transformation remains elusive. In this study, we probe the cellular and molecular mechanisms of RAS-mediated transformation, by profiling two human ovarian epithelial cell lines. One cell line was immortalized with SV40 T/t antigens and the human catalytic subunit of telomerase (T29), while the second cell line was transformed with an additional oncogenic rasV12 allele (T29H). In total, 32 proteins associated with RAS-mediated transformation have been identified using peptide mass fingerprinting. These protein targets are involved in several cellular pathways, including metabolism, redox balance, calcium signaling, apoptosis, and cellular methylation. One such target, the 40 kDa procaspase 4 is significantly upregulated at the protein level in RAS-transformed T29H cells, related directly to signaling through MEK, but not PI3 kinase. Cellular caspase 4 activity is, however, suppressed in the T29H cells, suggesting that the maturation process of caspase 4 is abrogated in RAS-transformed T29H cells. Consistent with this notion, transformed T29H cells were less susceptible to the toxic effects of anti-Fas antibody than were immortalized, nontransformed T29 cells, associated with less activation of caspase 4. This study demonstrates that functional proteomic analysis of a genetically defined cancer model provides a powerful approach toward systematically identifying cellular targets associated with oncogenic transformation.
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Abbreviations
- 2-DE:
-
two-dimensional electrophoresis
- ACN:
-
acetonitrile
- HOSE:
-
human ovarian surface epithelia cells
- hTERT:
-
human telomerase catalytic subunit
- MALDI-TOF:
-
matrix-assisted laser desorption/ionization time-of-flight
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Acknowledgements
We are indebted to Dr Natalie Ahn (University of Colorado, Boulder) for her continuous support and insightful discussions. We thank Dr Tony Haag in the Biomedical Resource Facility (supported by NCI grant R24CA88317, NIEHS Center Grant ES06676 and N01-HV-28184) for mass spectrometry analysis. This work is supported by American Cancer Society Research Scholar Grant RSG-01-035-01-TBE and National Institute of Health Grant GM060170 to XC. JC is supported by an American Cancer Society Research Scholar Grant RSG-04-028-01-CCE.
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Young, T., Mei, F., Liu, J. et al. Proteomics analysis of H-RAS-mediated oncogenic transformation in a genetically defined human ovarian cancer model. Oncogene 24, 6174–6184 (2005). https://doi.org/10.1038/sj.onc.1208753
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DOI: https://doi.org/10.1038/sj.onc.1208753
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