114P - Expression analysis of tumorspheres from non-small cell lung cancer show significant differences in CSC-markers and signaling pathways

Date 15 April 2016
Event European Lung Cancer Conference 2016 (ELCC) 2016
Session Poster lunch
Topics Non-Small-Cell Lung Cancer, Early Stage
Translational Research
Presenter Alejandro Herreros Pomares
Citation Journal of Thoracic Oncology (2016) 11 (supplement 4): S57-S166. S1556-0864(16)X0004-4
Authors A. Herreros Pomares1, E. Munera Maravilla1, B. Lafuente2, S. Calabuig-Fariñas3, A. Blasco Cordellat4, R. Guijarro5, R. Farrás6, E. Jantus Lewintre7, C. Camps8
  • 1Molecular Oncology Laboratory, General University Hospital Research Foundation, 46014 - Valencia/ES
  • 2Molecular Oncology Laboratory, General University Hospital Research Foundation, Valencia/ES
  • 3Department Of Pathology, Universitat De València, Molecular Oncology Laboratory, General University Hospital Research Foundation, 46014 - Valencia/ES
  • 4Servicio De Oncologia Medica, Hospital General Universitario Valencia, 46018 - Valencia/ES
  • 5Department Of Thoracic Surgery, Hospital General Universitario Valencia, Valencia/ES
  • 6Oncogenic Signaling, Centro de Investigación Príncipe Felipe, Valencia/ES
  • 7Department Of Biotechnology, Universidad Politécnica De Valencia, Molecular Oncology Laboratory, General University Hospital Research Foundation, Valencia/ES
  • 8Department Of Medicine, Universitat De València, Department Of Medical Oncology, Hospital General Universitario Valencia, Molecular Oncology Laboratory, General University Hospital Research Foundation, Valencia/ES

Abstract

Background

Despite the advances in the molecular characterization of lung cancer it remains as the leading cause of cancer death worldwide. Cancer stem cells (CSCs) are small subpopulations of stem-like cells with self-renewal and differentiation properties that constitute a promising target, but remain largely unknown. The aim of this study was to isolate and characterize gene expression of CSCs from lung cancer cell-lines and tumor-tissue obtained from resectable NSCLC patients.

Methods

This study was performed on cells from NSCLC tumor samples and cell lines (H1650, H1993, A549 and PC9) grown in monolayer and as spheroids. The expression of: CSC-markers (EPCAM1, ALDH1A1, CD166, ABCG2, CD44); pluripotency (KLF4, OCT4, NANOG, SOX2, MYC, CCND1); cell cycle (CDKN1A, CDKN2A, MDM2, WEE1); invasiveness (CDH1, CEACAM5, VIM, MMP2, MMP9); Notch pathway (NOTCH1, NOTCH2, NOTCH3, HEY1); Wnt pathway (CTNBB1, WNT1, WNT5A, DKK1, FZD7) and Hedgehog pathway (SMO, PTCH1, SHH, GLI1) were analyzed by RTqPCR. ACTB, CDKN1B and GUSB were used as endogenous controls for relative expression calculation.

Results

Lung tumorspheres had increased expression of EPCAM1, CD44, ALDH1A1 and CDKN1A (p = 0.028, p = 0.021, p = 0.043 and p = 0.021, respectively) when compared to their paired-adherent cells. Regarding the expression of Notch-pathway genes, NOTCH1 and NOTCH2 showed higher expression in tumorspheres (p = 0.028 and p = 0.038, respectively) and NOTCH3 also showed the same tendency. We found higher expression levels of CTNBB1 (Wnt pathway) (p = 0.008) in lungspheres whereas the activator of the non-canonical Wnt pathway, WNT5A, tended to be less expressed in spheroids compared to adherent culture cells. Our results show that SMO was underexpressed (p = 0.028) in tumorspheres, whereas no significant differences were found in other analyzed genes.

Conclusions

Lung spheroids from cancer cell lines and primary tumors showed increased levels of CSC-markers. Genes related to Notch and Wnt were found to be more expressed in tumorspheres, suggesting that these pathways as interesting lung-CSC targets.

Clinical trial identification

Legal entity responsible for the study

Fundación para la Investigación del Hospital General Universitario de Valencia

Funding

Supported partly by grants RD12/0036/0025 from RTICC, and PI12–02838/ PI15–0753 from ISCIII

Disclosure

All authors have declared no conflicts of interest.