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Rescue of glandular dysmorphogenesis in PTEN-deficient colorectal cancer epithelium by PPARγ-targeted therapy

Oncogene volume 32, pages 1305–1315 (2013)Cite this article

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Abstract

Disruption of glandular architecture associates with poor clinical outcome in high-grade colorectal cancer (CRC). Phosphatase and tensin homolog deleted on chromosome ten (PTEN) regulates morphogenic growth of benign MDCK (Madin Darby Canine Kidney) cells through effects on the Rho-like GTPase cdc42 (cell division cycle 42). This study investigates PTEN-dependent morphogenesis in a CRC model. Stable short hairpin RNA knockdown of PTEN in Caco-2 cells influenced expression or localization of cdc42 guanine nucleotide exchange factors and inhibited cdc42 activation. Parental Caco-2 cells formed regular hollow gland-like structures (glands) with a single central lumen, in three-dimensional (3D) cultures. Conversely, PTEN-deficient Caco-2 ShPTEN cells formed irregular glands with multiple abnormal lumens as well as intra- and/or intercellular vacuoles evocative of the high-grade CRC phenotype. Effects of targeted treatment were investigated. Phosphatidinylinositol 3-kinase (PI3K) modulating treatment did not affect gland morphogenesis but did influence gland number, gland size and/or cell size within glands. As PTEN may be regulated by the nuclear receptor peroxisome proliferator-activated receptor-γ (PPARγ), cultures were treated with the PPARγ ligand rosiglitazone. This treatment enhanced PTEN expression, cdc42 activation and rescued dysmorphogenesis by restoring single lumen formation in Caco-2 ShPTEN glands. Rosiglitazone effects on cdc42 activation and Caco-2 ShPTEN gland development were attenuated by cotreatment with GW9662, a PPARγ antagonist. Taken together, these studies show PTEN–cdc42 regulation of lumen formation in a 3D model of human CRC glandular morphogenesis. Treatment by the PPARγ ligand rosiglitazone, but not PI3K modulators, rescued colorectal glandular dysmorphogenesis of PTEN deficiency.

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Abbreviations

aPKC:

atypical protein kinase C

CA:

constitutively active

cdc42:

Cell division cycle 42

DN:

dominant negative

EGF:

epidermal growth factor

EV:

empty vector

GEFs:

Guanine nucleotide exchange factors

IGF:

insulin-like growth factor

PI3K:

Phosphatidylinositol 3-kinase

PtdIns:

Phosphatidylinositol

PTEN:

phosphatase and tensin homolog deleted on chromosome ten

Ser:

serine

ShRNA:

Short hairpin RNA

siRNA:

small interfering RNA

Thr:

threonine

wt:

wild type.

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Acknowledgements

We gratefully acknowledge the support of the Wellcome Trust (WT081232MA) and the Department for Education and Learning, Northern Ireland for financial support. We are greatly indebted to Dr T Waldman, Georgetown University, USA, for supply of PTEN+/+ HCT116 and PTEN −/−HCT116 cells, to Dr Pietro DiCamilli, Howard Hughes Medical Institute, Yale, for his generous provision of anti-human Tuba antibody, to Dr Estelle McLean for technical help, to Dr James Murray for the gift of PI-103 and to S Church for assistance with all imaging.

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  1. I Jagan, A Fatehullah and R K Deevi: These authors contributed equally to this study.

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  1. Centre for Cancer Research and Cell Biology, Queen’s University of Belfast, Belfast, UK

    I Jagan, A Fatehullah, R K Deevi, V Bingham & F C Campbell

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Correspondence to F C Campbell.

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Jagan, I., Fatehullah, A., Deevi, R. et al. Rescue of glandular dysmorphogenesis in PTEN-deficient colorectal cancer epithelium by PPARγ-targeted therapy. Oncogene 32, 1305–1315 (2013). https://doi.org/10.1038/onc.2012.140

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