Amplification of the Driving Oncogene, KRAS or BRAF, Underpins Acquired Resistance to MEK1/2 Inhibitors in Colorectal Cancer Cells
Annette S. Little,* Kathryn Balmanno, Matthew J. Sale, Scott Newman, Jonathan R. Dry, Mark Hampson, Paul A. W. Edwards, Paul D. Smith, Simon J. Cook*
*To whom correspondence should be addressed. E-mail:(A.S.L.); (S.J.C.).
This PDF file includes:
- Fig. S1. Growth curves of HCT116 and H6244-R1 or COLO205 and C6244-R1 cells and growth factor–independent proliferation.
- Fig. S2. AZD6244-resistant cells are cross-resistant to other MEK inhibitors.
- Fig. S3. AZD6244-resistant cells are not cross-resistant to cytotoxic drugs.
- Fig. S4. Kinetics of emergence of AZD6244 resistance.
- Fig. S5. Genomic PCR demonstrates amplification of BRAF and KRAS but not cyclin D1 in C6244-R1 and H6244-R1 cells.
- Fig. S6. BRAF and KRAS mutation analysis.
- Fig. S7. H6244-R1 cells show increased PI3K-PKB pathway signaling, but C6244-R1 cells do not.
- Fig. S8. Heat map of differential expression of a gene expression signature indicative of RAS activation in HCT116 versus H6244-R1 cells and COLO205 versus C6244-R1 cells.
- Fig. S9. Resistance to AZD6244 in C6244-R1 cells is reverted upon prolonged withdrawal of the drug.
- Fig. S10. Different methods of generating AZD6244-resistant cells give rise to a common mechanism of resistance.
- Fig. S11. HCT116 cells rendered resistant to PD184352 (H184-R) also show increased abundance of KRAS.
- Table S1. Summary of primers and probes used for the BRAF and KRAS qPCR assays.
- Table S2. Summary of primers used in the mutation screen to amplify the required regions for sequencing.
- Table S3. Summary of genetic background of cell lines used.
- Table S4. Weighting values relative to the performance of genes for the "MEK functional activation" and "Compensatory resistance" gene signatures specifically within colorectal cancer (CRC) samples.
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Citation: A. S. Little, K. Balmanno, M. J. Sale, S. Newman, J. R. Dry, M. Hampson, P. A. W. Edwards, P. D. Smith, S. J. Cook, Amplification of the Driving Oncogene, KRAS or BRAF, Underpins Acquired Resistance to MEK1/2 Inhibitors in Colorectal Cancer Cells. Sci. Signal. 4, ra17 (2011).