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Alpha2beta1 integrin regulates lineage commitment in multipotent human colorectal cancer cells.

Kirkland SC, Ying H - J. Biol. Chem. (2008)

Bottom Line: Function-blocking antibodies to alpha2 integrin also blocked both HRA-19 endocrine lineage commitment and enterocytic differentiation by Caco-2 human colon cancer cells; both effects being abrogated by the MEK inhibitor, PD98059, suggesting a role for ERK signaling in alpha2-mediated regulation of colorectal cancer cell differentiation.To further explore the role of alpha2 integrin in multilineage differentiation, we established multipotent cells expressing high levels of wild-type alpha2 integrin or a non-signaling chimeric alpha2 integrin.Overexpression of wild-type alpha2 integrin in HRA-19 cells significantly enhanced endocrine and mucous lineage commitment, while cells expressing the non-signaling chimeric alpha2 integrin had negligible ability for either endocrine or mucous lineage commitment.

View Article: PubMed Central - PubMed

Affiliation: Department of Histopathology, Faculty of Medicine, Imperial College London, London W12 ONN, United Kingdom. s.kirkland@imperial.ac.uk

ABSTRACT
The human colorectal epithelium is maintained by multipotent stem cells that give rise to absorptive, mucous, and endocrine lineages. Recent evidence suggests that human colorectal cancers are likewise maintained by a minority population of so-called cancer stem cells. We have previously established a human colorectal cancer cell line with multipotent characteristics (HRA-19) and developed a serum-free medium that induces endocrine, mucous and absorptive lineage commitment by HRA-19 cells in vitro. In this study, we investigate the role of the beta1 integrin family of cell surface extracellular matrix receptors in multilineage differentiation by these multipotent human colorectal cancer cells. We show that endocrine and mucous lineage commitment is blocked in the presence of function-blocking antibodies to beta1 integrin. Function-blocking antibodies to alpha2 integrin also blocked both HRA-19 endocrine lineage commitment and enterocytic differentiation by Caco-2 human colon cancer cells; both effects being abrogated by the MEK inhibitor, PD98059, suggesting a role for ERK signaling in alpha2-mediated regulation of colorectal cancer cell differentiation. To further explore the role of alpha2 integrin in multilineage differentiation, we established multipotent cells expressing high levels of wild-type alpha2 integrin or a non-signaling chimeric alpha2 integrin. Overexpression of wild-type alpha2 integrin in HRA-19 cells significantly enhanced endocrine and mucous lineage commitment, while cells expressing the non-signaling chimeric alpha2 integrin had negligible ability for either endocrine or mucous lineage commitment. This study indicates that the collagen receptor alpha2beta1 integrin is a regulator of cell fate in human multipotent colorectal cancer cells.

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α2β1 integrin is a collagen receptor in HRA-19, human colorectal cancer cells. Equal numbers of HRA-19 cells were seeded into wells coated with either human collagen I or IV and allowed to attach for 2 h at 37 °C. Attached cell number was measured using crystal violet staining and measurement at 595 nm: α1, 2 mg/ml; α2, 0.37 mg/ml; α3, 1.25 mg/ml; α5, 2 mg/ml; α6, 0.25 mg/ml; β1, 2253 5 mg/ml. Data are the mean ± S.D. (n = 3) **, p < 0.001; *, p < 0.005. Results are representative of a series of independent experiments performed on collagen I and collagen IV always including control wells and a range of antibodies; α1 (two experiments), α2 (four experiments), α3 (three experiments),α5 (three experiments), α6 (two experiments), β1 (five experiments).
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fig4: α2β1 integrin is a collagen receptor in HRA-19, human colorectal cancer cells. Equal numbers of HRA-19 cells were seeded into wells coated with either human collagen I or IV and allowed to attach for 2 h at 37 °C. Attached cell number was measured using crystal violet staining and measurement at 595 nm: α1, 2 mg/ml; α2, 0.37 mg/ml; α3, 1.25 mg/ml; α5, 2 mg/ml; α6, 0.25 mg/ml; β1, 2253 5 mg/ml. Data are the mean ± S.D. (n = 3) **, p < 0.001; *, p < 0.005. Results are representative of a series of independent experiments performed on collagen I and collagen IV always including control wells and a range of antibodies; α1 (two experiments), α2 (four experiments), α3 (three experiments),α5 (three experiments), α6 (two experiments), β1 (five experiments).

Mentions: α2β1 Integrin Is a Collagen Receptor in HRA-19 Cells—α2β1 is a major collagen receptor (37) in many cell types. Cell adhesion experiments were used to establish whether α2β1-mediated collagen binding in HRA-19 cells. Attachment to collagen I and IV was blocked by antibodies to β1 and α2 integrin (Fig. 4), indicating that α2β1 integrin is a receptor for both collagen I and IV in HRA-19 cells. Antibodies recognizing other α chains did not significantly reduce cell binding to either collagen I or collagen IV (Fig. 4).


Alpha2beta1 integrin regulates lineage commitment in multipotent human colorectal cancer cells.

Kirkland SC, Ying H - J. Biol. Chem. (2008)

α2β1 integrin is a collagen receptor in HRA-19, human colorectal cancer cells. Equal numbers of HRA-19 cells were seeded into wells coated with either human collagen I or IV and allowed to attach for 2 h at 37 °C. Attached cell number was measured using crystal violet staining and measurement at 595 nm: α1, 2 mg/ml; α2, 0.37 mg/ml; α3, 1.25 mg/ml; α5, 2 mg/ml; α6, 0.25 mg/ml; β1, 2253 5 mg/ml. Data are the mean ± S.D. (n = 3) **, p < 0.001; *, p < 0.005. Results are representative of a series of independent experiments performed on collagen I and collagen IV always including control wells and a range of antibodies; α1 (two experiments), α2 (four experiments), α3 (three experiments),α5 (three experiments), α6 (two experiments), β1 (five experiments).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2562061&req=5

fig4: α2β1 integrin is a collagen receptor in HRA-19, human colorectal cancer cells. Equal numbers of HRA-19 cells were seeded into wells coated with either human collagen I or IV and allowed to attach for 2 h at 37 °C. Attached cell number was measured using crystal violet staining and measurement at 595 nm: α1, 2 mg/ml; α2, 0.37 mg/ml; α3, 1.25 mg/ml; α5, 2 mg/ml; α6, 0.25 mg/ml; β1, 2253 5 mg/ml. Data are the mean ± S.D. (n = 3) **, p < 0.001; *, p < 0.005. Results are representative of a series of independent experiments performed on collagen I and collagen IV always including control wells and a range of antibodies; α1 (two experiments), α2 (four experiments), α3 (three experiments),α5 (three experiments), α6 (two experiments), β1 (five experiments).
Mentions: α2β1 Integrin Is a Collagen Receptor in HRA-19 Cells—α2β1 is a major collagen receptor (37) in many cell types. Cell adhesion experiments were used to establish whether α2β1-mediated collagen binding in HRA-19 cells. Attachment to collagen I and IV was blocked by antibodies to β1 and α2 integrin (Fig. 4), indicating that α2β1 integrin is a receptor for both collagen I and IV in HRA-19 cells. Antibodies recognizing other α chains did not significantly reduce cell binding to either collagen I or collagen IV (Fig. 4).

Bottom Line: Function-blocking antibodies to alpha2 integrin also blocked both HRA-19 endocrine lineage commitment and enterocytic differentiation by Caco-2 human colon cancer cells; both effects being abrogated by the MEK inhibitor, PD98059, suggesting a role for ERK signaling in alpha2-mediated regulation of colorectal cancer cell differentiation.To further explore the role of alpha2 integrin in multilineage differentiation, we established multipotent cells expressing high levels of wild-type alpha2 integrin or a non-signaling chimeric alpha2 integrin.Overexpression of wild-type alpha2 integrin in HRA-19 cells significantly enhanced endocrine and mucous lineage commitment, while cells expressing the non-signaling chimeric alpha2 integrin had negligible ability for either endocrine or mucous lineage commitment.

View Article: PubMed Central - PubMed

Affiliation: Department of Histopathology, Faculty of Medicine, Imperial College London, London W12 ONN, United Kingdom. s.kirkland@imperial.ac.uk

ABSTRACT
The human colorectal epithelium is maintained by multipotent stem cells that give rise to absorptive, mucous, and endocrine lineages. Recent evidence suggests that human colorectal cancers are likewise maintained by a minority population of so-called cancer stem cells. We have previously established a human colorectal cancer cell line with multipotent characteristics (HRA-19) and developed a serum-free medium that induces endocrine, mucous and absorptive lineage commitment by HRA-19 cells in vitro. In this study, we investigate the role of the beta1 integrin family of cell surface extracellular matrix receptors in multilineage differentiation by these multipotent human colorectal cancer cells. We show that endocrine and mucous lineage commitment is blocked in the presence of function-blocking antibodies to beta1 integrin. Function-blocking antibodies to alpha2 integrin also blocked both HRA-19 endocrine lineage commitment and enterocytic differentiation by Caco-2 human colon cancer cells; both effects being abrogated by the MEK inhibitor, PD98059, suggesting a role for ERK signaling in alpha2-mediated regulation of colorectal cancer cell differentiation. To further explore the role of alpha2 integrin in multilineage differentiation, we established multipotent cells expressing high levels of wild-type alpha2 integrin or a non-signaling chimeric alpha2 integrin. Overexpression of wild-type alpha2 integrin in HRA-19 cells significantly enhanced endocrine and mucous lineage commitment, while cells expressing the non-signaling chimeric alpha2 integrin had negligible ability for either endocrine or mucous lineage commitment. This study indicates that the collagen receptor alpha2beta1 integrin is a regulator of cell fate in human multipotent colorectal cancer cells.

Show MeSH
Related in: MedlinePlus