212P - Claudin-2: a new regulator of cancer stem cell self-renewal in colorectal cancer

Date 17 December 2016
Event ESMO Asia 2016 Congress
Session Poster lunch
Topics Colon and Rectal Cancer
Rectal Cancer
Translational Research
Presenter Sophie Paquet-Fifield
Citation Annals of Oncology (2016) 27 (suppl_9): ix53-ix67. 10.1093/annonc/mdw581
Authors S. Paquet-Fifield1, S.L. Koh2, L. Cheng3, L.M. Failla2, F. Hollande2
  • 1Pathology, The University of Melbourne, 3010 - Melbourne/AU
  • 2Pathology, The University of Melburne, 3010 - Melbourne/AU
  • 3Department Of Biochemistry And Genetics, La Trobe Institute for Molecular Science, 3083 - Melbourne/AU



Colorectal cancer (CRC) represents the third most lethal cancer worldwide. Fatal outcome often results from a recurrence of metastatic tumours, and a subpopulation of cancer cells called Cancer Stem Cells (CSCs) is thought to be instrumental for this process. Although the overexpression of the tight junction protein claudin-2 correlates with cancer progression in human CRC, and its expression in vitro enhances cell proliferation and colony formation and reduces chemosensitivity of CRC cells, the role of claudin-2 in regulating colorectal CSCs remains unknown.


We experimentally overexpressed or down-regulated claudin-2 expression in unique patient-derived cells from primary colorectal tumours and liver metastases, as well as in several classical CRC cell lines, depending on their endogenous level of claudin-2 expression. We performed stem cell assays in vitro (Extreme Limiting Dilution Assay, colonosphere formation assays) and xenograft experiments in vivo to demonstrate that claudin-2 promotes self-renewal of CSCs. We also performed qRT-PCR analysis of stage II/III CRC tumours post-5-FU-based adjuvant chemotherapy, combined with data mining of two large genomic datasets of equivalent samples. Next Generation Sequencing was performed to further analyse miRNAs expression and pathways were analysed.


Claudin-2 expression correlates with higher probability of recurrence and poor prognosis in the clinic. Based on the quantification of stem cell activity (ALDH activity), we further show that claudin-2 regulates phenotypic transitions between CSCs and non-CSCs phenotypes. Finally, Next Generation Sequencing analyses comparing levels of miRNAs between the SC-like population expressing or not claudin-2, identified a list of 9 miRNAs directly involved in Wnt/beta-catenin, Stem Cell and Colorectal cancer signalling pathways. Further insights into the molecular regulation of CSCs in CRC by claudin-2 be discussed.


Our findings demonstrate for the first time that claudin-2 regulates the self-renewal tumour-initiating ability of colorectal CSCs. These new molecular insights are novel and important to lead the development of future therapeutic strategies targeting CSCs against post-treatment recurrence.

Clinical trial indentification

Legal entity responsible for the study

University of Melbourne


University of Melbourne


S. Paquet-Fifield: The presenter and all co-authors do not identify any financial interest in products or processes involved in their research. S.L. Koh: no financial interest. All other authors have declared no conflicts of interest.