Colon cancer cells adopt an invasive phenotype without mesenchymal transition in 3-D but not 2-D culture upon combined stimulation with EGF and crypt growth factorsReportar como inadecuado




Colon cancer cells adopt an invasive phenotype without mesenchymal transition in 3-D but not 2-D culture upon combined stimulation with EGF and crypt growth factors - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

BMC Cancer

, 13:221

Cell and molecular biology

Abstract

BackgroundThe intestinal crypt homeostasis is maintained by a combination of growth factors including Wnt, R-Spondin1, Noggin and the epidermal growth factor EGF. In human colorectal cancer, the Wnt pathway is constitutively activated through genetic and epigenetic alterations in as many as 11 genes encoding components of this crypt stem-cell maintenance mechanism. Although the proliferation of colon cancer cells does not require Wnt, it is possible that colon cancer cells can still respond to the crypt growth factors in the colonic microenvironment. A number of studies have shown that epithelial cells behave differently in 3-D versus 2-D cultures. Because the 3-D conditions more closely mimic the in vivo environment, we examined the effects of Wnt and other crypt growth factors on colon cancer cell growth in 3-D culture.

MethodsColon cancer cells were grown in 3-D matrigel supplemented with different combinations of crypt growth factors and colonies were examined for morphology and pathways.

ResultsWhen colon cancer cells were cultured in 3-D with EGF, they grew as round spheroid colonies. However, colon cancer cells also grew as flat, disc-like colonies when cultured with EGF plus Wnt, R-Spondin1 and Noggin. Disc colonies were found to have comparable levels of E-cadherin as the spheroid colonies, but showed decreased E-cadherin at the cell-matrix contact sites. Disc colonies also elaborated F-actin rich protrusions FRP at the cell-matrix edge, reminiscent of an invasive phenotype but without the expression of vimentin. These E-cadherin and F-actin alterations were not induced by the four growth factors in 2-D culture. Formation of the disc colonies was inhibited by the knockdown of β-catenin and by protein kinase inhibitors such as gefitinib, imatinib and MK-2206. Furthermore, withdrawal of the crypt growth factors was able to revert the disc colonies to spheroid growth, showing that the invasive phenotype was reversible dependent on the availability of growth factors.

ConclusionsThese findings show that colon cancer cells remain responsive to the growth factors in the crypt microenvironment and can be induced to undergo morphological transformation in the more physiologically relevant 3-D culture.

AbbreviationsEGFEpidermal Growth Factor

MMPMatrix-Metallo Proteases

FAKFocal Adhesion Kinase

FRPF-actin Rich Protrusions

ECMExtra Cellular Matrix

TCGAThe Cancer Genome Atlas

CRCColo-Rectal Cancer

TGF-βTransforming Growth Factor

BMPBone Morphogenetic Protein

RTKReceptor Tyrosine Kinase

RR-Spondin1

NNoggin

EEGF

WWnt3a

RNEWR-Spondin1, Noggin, EGF, Wnt3a.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2407-13-221 contains supplementary material, which is available to authorized users.

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Autor: Kirsten Ludwig - Edison S Tse - Jean YJ Wang

Fuente: https://link.springer.com/







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