1. PAR2-dependent activation of GSK3 beta regulates the survival of colon stem/progenitor cells
- Author
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Emmanuel Mas, Emilie d'Aldebert, Raoudha Mezghani-Jarraya, Bailey Zwarycz, Nathalie Vergnolle, Claire Racaud-Sultan, Nicolas Cenac, Delphine Bonnet, Scott T. Magness, Corinne Rolland, Muriel Quaranta, Laurianne Van Landeghem, Audrey Ferrand, Sokchea Khou, Chrystelle Bonnart, Imen Nasri, ProdInra, Archive Ouverte, Institut de Recherche en Santé Digestive (IRSD ), Institut National de la Recherche Agronomique (INRA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM), Service Médecine Interne et immunologie clinique [CHU Toulouse], Pôle Maladies de l'appareil digestif [CHU Toulouse], Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Departments of Medicine and Cell Biology & Physiology, University of North Carolina [Chapel Hill] (UNC), University of North Carolina System (UNC)-University of North Carolina System (UNC), Laboratoire de Chimie des Substances Naturelles, Faculté des Sciences de Sfax, Université de Sfax - University of Sfax, Service de Gastroentérologie et pancréatologie [CHU Toulouse], Association de Recherche contre le Cancer(www.fondation-arc.org, no. SFI20121205496, N. Vergnolle), Ligue Ré-gionale contre le Cancer (www.ligue-cancer.net, 2013 Aude Comity, C.Racaud-Sultan), the Institut National du Cancer/Aviesan (www.e-cancer.fr, no.P029564, D. Bonnet), the Programme Hubert Curien Utique cooperation(www.egide.asso.fr, no. CMCU12G0821, R. Mezghani-Jarraya, C. Racaud-Sultan), the Région Midi-Pyrénées (www.midipyrenees.fr, no. 12050835, Ed’Aldebert), European Research Council Starting Grant (erc.europa.eu,ERC_2012_StG_20111109, N. Vergnolle), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Service de Médecine Interne, Fédération Digestive, CHU Toulouse [Toulouse]-Hôpital Purpan [Toulouse], CHU Toulouse [Toulouse], Service de Gastroentérologie, Hépatologie et Nutrition, Centre Hospitalier Universitaire de Toulouse, U913 Neuropathies du système nerveux entérique et pathologies digestives, and Université de Nantes (UN)
- Subjects
0301 basic medicine ,Male ,Physiology ,Colorectal cancer ,0302 clinical medicine ,Tumor Microenvironment ,Protease-activated receptor ,glycogen synthase kinase-3 beta ,Protein Phosphatase 2 ,Phosphorylation ,Stem Cell Niche ,colorectal cancer ,protease-activated receptor ,inflammation ,spheroid ,Receptor ,rho-Associated Kinases ,Arrestin ,[SDV.MHEP] Life Sciences [q-bio]/Human health and pathology ,Stem Cells ,Gastroenterology ,[SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism ,3. Good health ,Cell biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Neoplastic Stem Cells ,Endocrinologie et métabolisme ,RNA Interference ,medicine.symptom ,Signal Transduction ,Cell Survival ,Colon ,Stem Cells, Tissue Engineering, Development, and Cancer ,Inflammation ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,macromolecular substances ,Biology ,Transfection ,03 medical and health sciences ,Physiology (medical) ,Spheroids, Cellular ,medicine ,Animals ,Humans ,Receptor, PAR-2 ,Progenitor cell ,GSK3B ,[SDV.BC] Life Sciences [q-bio]/Cellular Biology ,Progenitor ,Cell Proliferation ,Endocrinology and metabolism ,Glycogen Synthase Kinase 3 beta ,Hepatology ,Epithelial Cells ,medicine.disease ,Epithelium ,Enzyme Activation ,Mice, Inbred C57BL ,030104 developmental biology ,Caco-2 Cells ,[SDV.MHEP]Life Sciences [q-bio]/Human health and pathology - Abstract
Protease-activated receptors PAR1 and PAR2 play an important role in the control of epithelial cell proliferation and migration. However, the survival of normal and tumor intestinal stem/progenitor cells promoted by proinflammatory mediators may be critical in oncogenesis. The glycogen synthase kinase-3β (GSK3β) pathway is overactivated in colon cancer cells and promotes their survival and drug resistance. We thus aimed to determine PAR1 and PAR2 effects on normal and tumor intestinal stem/progenitor cells and whether they involved GSK3β. First, PAR1 and PAR2 were identified in colon stem/progenitor cells by immunofluorescence. In three-dimensional cultures of murine crypt units or single tumor Caco-2 cells, PAR2 activation decreased numbers and size of normal or cancerous spheroids, and PAR2-deficient spheroids showed increased proliferation, indicating that PAR2 represses proliferation. PAR2-stimulated normal cells were more resistant to stress (serum starvation or spheroid passaging), suggesting prosurvival effects of PAR2. Accordingly, active caspase-3 was strongly increased in PAR2-deficient normal spheroids. PAR2 but not PAR1 triggered GSK3β activation through serine-9 dephosphorylation in normal and tumor cells. The PAR2-triggered GSK3β activation implicates an arrestin/PP2A/GSK3β complex that is dependent on the Rho kinase activity. Loss of PAR2 was associated with high levels of GSK3β nonactive form, strengthening the role of PAR2 in GSK3β activation. GSK3 pharmacological inhibition impaired the survival of PAR2-stimulated spheroids and serum-starved cells. Altogether our data identify PAR2/GSK3β as a novel pathway that plays a critical role in the regulation of stem/progenitor cell survival and proliferation in normal colon crypts and colon cancer.
- Published
- 2016
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