Basic—Alimentary TractExperimentally Derived Metastasis Gene Expression Profile Predicts Recurrence and Death in Patients With Colon Cancer
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Cell Culture and Mouse Model
MC-38 mouse adenocarcinoma cells were obtained from the American Type Culture Collection (Manassas, VA).9 MC-38 cells were transfected with firefly luciferase gene (pGL3 basic; Promega, Madison, WI) and selected (G418; Invitrogen, Carlsbad, CA). To enrich for invasive cells, 7.5 × 105 cells were seeded onto 6-well, 8.0-micron pore transwell polycarbonate membrane inserts (Costar, Cambridge, MA) coated with 2.5 mg/mL matrigel, and incubated with serum-free Dulbecco's modified Eagle's medium in
Development of an Immunocompetent Mouse Model of Colon Cancer Metastasis
Tumors are a heterogeneous mixture of cells with varied invasive and metastatic potentials. Therefore, we used a conventional invasion assay to enrich for a subpopulation of highly invasive MC-38 mouse colon cancer cells (Figure 1A, MC-38inv). After 6 serial passages through matrigel, MC-38inv cells were more invasive than MC-38 parental cells both in vitro and in vivo in a tail vein injection assay (Figure 1B and Supplementary Table 1; P < .001). Lung tumors derived from MC-38inv cells were
Discussion
In the present study, the biology of colon cancer metastasis was modeled in immunocompetent mice to develop a gene expression classifier that discriminates recurrence and survival outcomes in human patients with colon cancer. Patients with stage II and stage III primary colon cancers that reflected the recurrence-associated gene expression pattern were at greater relative risk of recurrence than patients who did not (HR = 13.1 and 4.7, respectively). This gene expression profile, tested with a
Acknowledgments
The authors thank James Goldenring, MD, PhD, Eric Neilson, MD, and Vivien Siegel, PhD for critically reviewing the manuscript in advance of submission. We thank Ramona Deal for derivation of luciferase-expressing MC-38 parental cells. We thank the Vanderbilt Microarray Shared Resource. We gratefully acknowledge the generous philanthropic support of the Ingram family. J.J.S. gratefully acknowledges support from the Society of University Surgeons-Ethicon Scholarship Award, the Vanderbilt Clinical
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Cited by (0)
The current address for Fei Wu is Bureau of Communicable Disease and Prevention, Missouri Department of Health and Senior Services, 930 Wildwood, PO Box 570, Jefferson City, Missouri 65102. The current address for Carl Schmidt is The Ohio State University Medical Center, College of Medicine, Division of Surgical Oncology, N-924 Doan Hall, 410 West 10th Avenue, Columbus, OH 43210-1240.
GEO accession number is GSE17538. Complete Minimum information about a microarray experiment (MIAME)-compliant datasets for analysis is available (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE17538).
Conflicts of interest The authors disclose no conflicts.
Funding This work was supported by the following grants from the National Institutes of Health: CA69457, DK52334, CA068485, and CA077839 (R.D.B.); TL1 RR024978 and CA106183 (J.J.S.); CA46413, CA95103, and CA084239 (R.J.C.); CA112215 (T.J.Y.); and CA126588 and CA128323 (N.G.D.). Other sources of funding include the Society of University Surgeons-Ethicon Scholarship Award (J.J.S.).
J. Joshua Smith, Natasha G. Deane, Fei Wu, and R. Daniel Beauchamp contributed equally to this work.