Identification of mutant K-Ras-dependent phenotypes using a panel of isogenic cell lines
Open accessAbstract
To assess the consequences of endogenous mutant K-Ras, we analyzed the signaling and biological properties of a small panel of isogenic cell lines. These include the cancer cell lines DLD1, HCT116, and Hec1A, in which either the WT or mutant K- ras allele has been disrupted, and SW48 colorectal cancer cells and human mammary epithelial cells in which a single copy of mutant K- ras was introduced at its endogenous genomic locus. We find that single copy mutant K-Ras causes surprisingly modest activation of downstream signaling to ERK and Akt. In contrast, a negative feedback signaling loop to EGFR and N-Ras occurs in some, but not all, of these cell lines. Mutant K-Ras also had relatively minor effects on cell proliferation and cell migration but more dramatic effects on cell transformation as assessed by growth in soft agar. Surprisingly, knock-out of the wild type K- ras allele consistently increased growth in soft agar, suggesting tumor-suppressive properties of this gene under these conditions. Finally, we examined the effects of single copy mutant K-Ras on global gene expression. Although transcriptional programs triggered by mutant K-Ras were generally quite distinct in the different cell lines, there was a small number of genes that were consistently overexpressed, and these could be used to monitor K-Ras inhibition in a panel of human tumor cell lines. We conclude that there are conserved components of mutant K-Ras signaling and phenotypes but that many depend on cell context and environmental cues. Background: Many studies analyzing K-Ras function rely on overexpression. Results: Knock-out and knock-in of endogenous mutant K- ras have modest effects on downstream signaling but strong effects on gene expression and transformation. Conclusion: Genomic manipulation allows physiological determination of WT and mutant K-Ras consequences. Significance: Gene expression patterns can be used to monitor inhibition of mutant K-Ras.
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Not all KRAS-mutant tumor cells are dependent on KRAS.
“Early evidence suggested that not all KRAS-mutant tumor cells are dependent on KRAS (18, 19)”
From New Horizons in KRAS-Mutant Lung Cancer: Dawn After Darkness (2019) · cites this paper as [19]
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- Authors
- Vartanian, S., Bentley, C., Brauer, M. J., Li, L., Shirasawa, S., Sasazuki, T., Kim, J.-S., Haverty, P., Stawiski, E., Modrusan, Z., Waldman, T., & Stokoe, D.
- Affiliations
- Genentech (US)
- Fukuoka University (JP)
- Kyushu University (JP)
- Georgetown University (US)
- Corresponding author
- Stokoe, David
- ORCID
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2013-01
- Volume
- 288
- Issue
- 4
- Pages
- 2403-2413
- ISSN
- 0021-9258 (ISSN-L)
- PMC
- PMC3554910
- OpenAlex
- W2010172360
- Cited by
- 65
- References
- 44
- Citations by year
- 2025: 3 · 2022: 8 · 2021: 9 · 2020: 4 · 2019: 7 · 2018: 8 · 2017: 4 · 2016: 4 · 2015: 8 · 2014: 7 · 2013: 3
- FWCI
- 2.31
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Genentech
- Japan Society for the Promotion of Science (21390102)
- National Cancer Institute (P30CA016672)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Tyrosine Phosphatases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Melanoma and MAPK Pathways
- Keywords
- mutant KRAS, isogenic cell lines, ERK signaling, Akt signaling, NRAS, cell transformation, soft agar growth, tumor suppressor function, gene expression profiling, colorectal cancer, mammary epithelial cells
- MeSH terms
- Cell Line, Cell Movement, Heterozygote, Homozygote, Humans, Mutation, Oncogenes, Phenotype, Genes, ras, Transcription, Genetic, Signal Transduction, Gene Expression Regulation, Neoplastic, ras Proteins, Gene Expression Profiling, RNA, Small Interfering, Cell Line, Tumor, Cell Proliferation
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