KIT Signaling Promotes Growth of Colon Xenograft Tumors in Mice and Is Up-Regulated in a Subset of Human Colon Cancers

Evan C. Chen, Taylor A. Karl, Tomer Kalisky, Santosh K. Gupta, Catherine A. O'Brien, Teri A. Longacre, Matt Van De Rijn, Stephen R. Quake, Michael F. Clarke, Michael E. Rothenberg

Research output: Contribution to journalArticlepeer-review

37 Scopus citations

Abstract

Background & Aims Receptor tyrosine kinase (RTK) inhibitors have advanced colon cancer treatment. We investigated the role of the RTK KIT in development of human colon cancer. Methods An array of 137 patient-derived colon tumors and their associated xenografts were analyzed by immunohistochemistry to measure levels of KIT and its ligand KITLG. KIT and/or KITLG was stably knocked down by expression of small hairpin RNAs from lentiviral vectors in DLD1, HT29, LS174T, and COLO320 DM colon cancer cell lines, and in UM-COLON#8 and POP77 xenografts; cells transduced with only vector were used as controls. Cells were analyzed by real-time quantitative reverse transcription polymerase chain reaction, single-cell gene expression analysis, flow cytometry, and immunohistochemical, immunoblot, and functional assays. Xenograft tumors were grown from control and KIT-knockdown DLD1 and UM-COLON#8 cells in immunocompromised mice and compared. Some mice were given the RTK inhibitor imatinib after injection of cancer cells; tumor growth was measured based on bioluminescence. We assessed tumorigenicity using limiting dilution analysis. Results KIT and KITLG were expressed heterogeneously by a subset of human colon tumors. Knockdown of KIT decreased proliferation of colon cancer cell lines and growth of xenograft tumors in mice compared with control cells. KIT knockdown cells had increased expression of enterocyte markers, decreased expression of cycling genes, and, unexpectedly, increased expression of LGR5 associated genes. No activating mutations in KIT were detected in DLD1, POP77, or UM-COLON#8 cells. However, KITLG-knockdown DLD1 cells formed smaller xenograft tumors than control cells. Gene expression analysis of single CD44+ cells indicated that KIT can promote growth via KITLG autocrine and/or paracrine signaling. Imatinib inhibited growth of KIT+ colon cancer organoids in culture and growth of xenograft tumors in mice. Cancer cells with endogenous KIT expression were more tumorigenic in mice. Conclusions KIT and KITLG are expressed by a subset of human colon tumors. KIT signaling promotes growth of colon cancer cells and organoids in culture and xenograft tumors in mice via its ligand, KITLG, in an autocrine or paracrine manner. Patients with KIT-expressing colon tumors can benefit from KIT RTK inhibitors.

Original languageEnglish
Pages (from-to)705e2-717.e2
JournalGastroenterology
Volume149
Issue number3
DOIs
StatePublished - 1 Sep 2015

Bibliographical note

Publisher Copyright:
© 2015 AGA Institute.

Funding

Funding This study was supported by National Institutes of Health (NIH), National Institute of Diabetes and Digestive and Kidney Diseases K08-DK097181 (MER), NIH S10 Shared Instrumentation Grant 1S10RR02933801, NIH National Institute of Neurological Disorders and Stroke NS069375 , Howard Hughes Medical Institute Medical Research Fellowship (ECC).

FundersFunder number
National Institutes of Health
Howard Hughes Medical Institute
National Institute of Diabetes and Digestive and Kidney DiseasesK08DK097181, 1S10RR02933801
National Institute of Neurological Disorders and StrokeP30NS069375

    Keywords

    • CD117
    • Colorectal Cancer
    • Enteroids
    • Stem Cell Factor

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