Chronic chemotherapeutic stress promotes evolution of stemness and WNT/beta-catenin signaling in colorectal cancer cells: implications for clinical use of WNT-signaling inhibitors

Abstract

// Meriam Ayadi 1, 2, 3 , Anaïs Bouygues 1, 2, 3 , Djamila Ouaret 1, 2, 3 , Nathalie Ferrand 1, 2, 3 , Salem Chouaib 4 , Jean-Paul Thiery 5, 6, 7 , Christian Muchardt 8 , Michèle Sabbah 1, 2, 3 , Annette K Larsen 1, 2, 3 1 Cancer Biology and Therapeutics, Centre de Recherche Saint-Antoine, Paris, France 2 Institut National de la Santé et de la Recherche Médicale (INSERM), Paris, France 3 Institut Universitaire de Cancérologie, Pierre et Marie Curie (UPMC) Sorbonne Universités, Paris, France 4 Institut National de la Santé et de la Recherche Médicale (INSERM), Gustave-Roussy, Villejuif, France 5 Institute of Molecular and Cell Biology, A*STAR (Agency for Science, Technology and Research), Singapore, Singapore. 6 Department of Biochemistry, School of Medicine, National University of Singapore, Singapore. 7 Cancer Science Institute, National University of Singapore, Singapore 8 Laboratory of Epigenetic Regulation, Centre National de la Recherche Scientifique (CNRS), Institut Pasteur, Paris, France Correspondence to: Annette K. Larsen, e-mail: annette.larsen@upmc.fr Keywords: colorectal cancer, chemotherapeutic stress, “stemness”, WNT/beta-catenin signaling, WNT inhibitors Received: January 05, 2015      Accepted: April 29, 2015      Published: May 11, 2015 ABSTRACT Most solid tumors contain a subfraction of cells with stem/progenitor cell features. Stem cells are naturally chemoresistant suggesting that chronic chemotherapeutic stress may select for cells with increased “stemness”. We carried out a comprehensive molecular and functional analysis of six independently selected colorectal cancer (CRC) cell lines with acquired resistance to three different chemotherapeutic agents derived from two distinct parental cell lines. Chronic drug exposure resulted in complex alterations of stem cell markers that could be classified into three categories: 1) one cell line, HT-29/5-FU, showed increased “stemness” and WNT-signaling, 2) three cell lines showed decreased expression of stem cell markers, decreased aldehyde dehydrogenase activity, attenuated WNT-signaling and lost the capacity to form colonospheres and 3) two cell lines displayed prominent expression of ABC transporters with a heterogeneous response for stem cell markers. While WNT-signaling could be attenuated in the HT-29/5-FU cells by the WNT-signaling inhibitors ICG-001 and PKF-118, this was not accompanied by any selective growth inhibitory effect suggesting that the cytotoxic activity of these compounds is not directly linked to WNT-signaling inhibition. We conclude that classical WNT-signaling inhibitors have toxic off-target activities that need to be addressed for clinical development.