CD133+ colon cancer cells are more interactive with the tumor microenvironment than CD133-cells

Celia Chao, J. Russ Carmical, Kirk L. Ives, Thomas Wood, Judith Aronson, Guillermo Gomez, Clarisse D. Djukom, Mark Hellmich

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Experimental data indicate that colorectal cancer cells with CD133+ expression exhibit enhanced tumorigenicity over CD133+-negative (CD133+) cells. We hypothesized that CD133+-positive (CD133+) cells, compared with CD133+, are more tumorigenic because they are more interactive with and responsive to their stromal microenvironment. Freshly dissected and dissociated cells from a primary colon cancer were separated into carcinoma-associated fibroblasts (CAF) and the epithelial cells; the latter were further separated into CD133+ and CD133+ cells using fluorescence-activated cell sorter. The CD133+ cells formed large tumors in non-obese diabetic-severe combined immunodeficient (NOD-SCID) mice, demonstrating the phenotypic cellular diversity of the original tumor, whereas CD133+ cells were unable to sustain significant growth. Affymetrix gene array analyses using t-test, fold-change and multiple test correction identified candidate genes that were differentially expressed between the CD133+ vs CD133+ cells. RT-PCR verified differences in expression for 30 of the 46 genes selected. Genes upregulated ( vs-cells) included CD133+ (9.3-fold) and CXCR4 (4-fold), integrin Β8 and fibroblast growth factor receptor 2. The CAF highly express the respective ligands: stromal-derived factor-1 (SDF-1), vitronectin and FGF family members, suggesting a reciprocal relationship between the CD133+ and CAF cells. SDF-1 caused an increase in intracellular calcium in cells expressing both CD133+ and CXCR4, confirming functional CXCR4. The CD133 +CXCR4 phenotype is increased to 32% when the cells are grown in suspension compared with only 9% when the cells were allowed to attach. In Matrigel 3-D culture, the CD133 +CXCR4 group treated with SDF-1 grew more colonies compared with vehicle, as well as significantly larger colony sizes of tumor spheres. These data demonstrate proof of principle that the enhanced tumorigenic potential of CD133+, compared with CD133+, cells is due to their increased ability to interact with their neighboring CAF.

Original languageEnglish (US)
Pages (from-to)420-436
Number of pages17
JournalLaboratory Investigation
Volume92
Issue number3
DOIs
StatePublished - Mar 2012

Fingerprint

Tumor Microenvironment
Colonic Neoplasms
Fibroblasts
Carcinoma
Genes
Receptor, Fibroblast Growth Factor, Type 2
Vitronectin
Fibroblast Growth Factor 1
Neoplasms
Aptitude
SCID Mice
Integrins

Keywords

  • CD133
  • colon cancer
  • CXCR4
  • tumor microenvironment

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Cell Biology
  • Molecular Biology

Cite this

CD133+ colon cancer cells are more interactive with the tumor microenvironment than CD133-cells. / Chao, Celia; Carmical, J. Russ; Ives, Kirk L.; Wood, Thomas; Aronson, Judith; Gomez, Guillermo; Djukom, Clarisse D.; Hellmich, Mark.

In: Laboratory Investigation, Vol. 92, No. 3, 03.2012, p. 420-436.

Research output: Contribution to journalArticle

Chao, Celia ; Carmical, J. Russ ; Ives, Kirk L. ; Wood, Thomas ; Aronson, Judith ; Gomez, Guillermo ; Djukom, Clarisse D. ; Hellmich, Mark. / CD133+ colon cancer cells are more interactive with the tumor microenvironment than CD133-cells. In: Laboratory Investigation. 2012 ; Vol. 92, No. 3. pp. 420-436.
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abstract = "Experimental data indicate that colorectal cancer cells with CD133+ expression exhibit enhanced tumorigenicity over CD133+-negative (CD133+) cells. We hypothesized that CD133+-positive (CD133+) cells, compared with CD133+, are more tumorigenic because they are more interactive with and responsive to their stromal microenvironment. Freshly dissected and dissociated cells from a primary colon cancer were separated into carcinoma-associated fibroblasts (CAF) and the epithelial cells; the latter were further separated into CD133+ and CD133+ cells using fluorescence-activated cell sorter. The CD133+ cells formed large tumors in non-obese diabetic-severe combined immunodeficient (NOD-SCID) mice, demonstrating the phenotypic cellular diversity of the original tumor, whereas CD133+ cells were unable to sustain significant growth. Affymetrix gene array analyses using t-test, fold-change and multiple test correction identified candidate genes that were differentially expressed between the CD133+ vs CD133+ cells. RT-PCR verified differences in expression for 30 of the 46 genes selected. Genes upregulated ( vs-cells) included CD133+ (9.3-fold) and CXCR4 (4-fold), integrin Β8 and fibroblast growth factor receptor 2. The CAF highly express the respective ligands: stromal-derived factor-1 (SDF-1), vitronectin and FGF family members, suggesting a reciprocal relationship between the CD133+ and CAF cells. SDF-1 caused an increase in intracellular calcium in cells expressing both CD133+ and CXCR4, confirming functional CXCR4. The CD133 +CXCR4 phenotype is increased to 32{\%} when the cells are grown in suspension compared with only 9{\%} when the cells were allowed to attach. In Matrigel 3-D culture, the CD133 +CXCR4 group treated with SDF-1 grew more colonies compared with vehicle, as well as significantly larger colony sizes of tumor spheres. These data demonstrate proof of principle that the enhanced tumorigenic potential of CD133+, compared with CD133+, cells is due to their increased ability to interact with their neighboring CAF.",
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AB - Experimental data indicate that colorectal cancer cells with CD133+ expression exhibit enhanced tumorigenicity over CD133+-negative (CD133+) cells. We hypothesized that CD133+-positive (CD133+) cells, compared with CD133+, are more tumorigenic because they are more interactive with and responsive to their stromal microenvironment. Freshly dissected and dissociated cells from a primary colon cancer were separated into carcinoma-associated fibroblasts (CAF) and the epithelial cells; the latter were further separated into CD133+ and CD133+ cells using fluorescence-activated cell sorter. The CD133+ cells formed large tumors in non-obese diabetic-severe combined immunodeficient (NOD-SCID) mice, demonstrating the phenotypic cellular diversity of the original tumor, whereas CD133+ cells were unable to sustain significant growth. Affymetrix gene array analyses using t-test, fold-change and multiple test correction identified candidate genes that were differentially expressed between the CD133+ vs CD133+ cells. RT-PCR verified differences in expression for 30 of the 46 genes selected. Genes upregulated ( vs-cells) included CD133+ (9.3-fold) and CXCR4 (4-fold), integrin Β8 and fibroblast growth factor receptor 2. The CAF highly express the respective ligands: stromal-derived factor-1 (SDF-1), vitronectin and FGF family members, suggesting a reciprocal relationship between the CD133+ and CAF cells. SDF-1 caused an increase in intracellular calcium in cells expressing both CD133+ and CXCR4, confirming functional CXCR4. The CD133 +CXCR4 phenotype is increased to 32% when the cells are grown in suspension compared with only 9% when the cells were allowed to attach. In Matrigel 3-D culture, the CD133 +CXCR4 group treated with SDF-1 grew more colonies compared with vehicle, as well as significantly larger colony sizes of tumor spheres. These data demonstrate proof of principle that the enhanced tumorigenic potential of CD133+, compared with CD133+, cells is due to their increased ability to interact with their neighboring CAF.

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