Author of

• 16751

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  P1.05 - Poster Session with Presenters Present (ID 457)

  • Event: WCLC 2016
  • Type: Poster Presenters Present
  • Track: Early Stage NSCLC
  • Presentations: 1
  • Moderators:
  • Coordinates: 12/05/2016, 14:30 - 15:45, Hall B (Poster Area)

  • 16765

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    P1.05-014 - Stemness Gene Expression Profile of Tumorspheres from Non-Small Cell Lung Cancer (ID 4748)

    14:30 - 14:30  |  Author(s): E. García Del Olmo
    • Abstract

    Background:
    Lung cancer features like chemoresistance, tumor progression or metastasis have consolidated lung cancer as the first cause of death cancer-related worldwide. Cancer stem cells (CSCs) are small subpopulations of stem-like cells that have been associated to these traits, constituting a promising target, but remaining largely unknown. In this study, we isolated CSCs from lung cancer cell lines and tumor tissue of resectable NSCLC patients using a sphere-forming assay and analyzed their gene expression profile.
    
    Methods:
    The investigation was carried out on cells from seven NSCLC tumor samples and six cell lines (H1650, H1993, H358, A549, PC9, H460) grown in monolayer and as spheroids. The expression of CSC-markers (CD133, EPCAM1, ALDH1A1, CD166, ABCG2, CD44, MUC1, BMI1, THY1), pluripotency promoters (KLF4, OCT4, NANOG, SOX2, MYC, CCND1), cell cycle regulators (CDKN1A, CDKN2A, MDM2, WEE1), invasiveness-related genes (CDH1, VIM, SNAI1, MMP2, MMP9, CEACAM5, ITGA2, ITGA6, ITGB1), Notch pathway (NOTCH1, NOTCH2, NOTCH3, JAG1, DLL1, DLL4, NUMB, HEY1, HES1), Wnt pathway (WNT1, WNT2, WNT3, WNT5A, CTNBB1, DKK1, FZD7) and Hedgehog pathway (SMO, PTCH1, SHH, GLI1) components were analyzed by quantitative real time PCR (RTqPCR). ACTB, CDKN1B and GUSB genes were used as housekeeping controls for the relative expression calculation.
    
    Results:
    Lungspheres showed significantly higher expression of the CSC-markers EPCAM1, CD44 and ALDH1A1 (p= 0.028, p= 0.021 and p= 0.043, respectively) and the quiescence promoter CDKN1A (p= 0.021) in comparison with their paired-monolayer cells. The epithelial to mesenquimal transition (EMT) inducer, SNAI1, as well as integrins ITGA2, ITGA6 and ITGB1 were overexpressed in tumorspheres (p= 0.011, p= 0.018, p= 0.016 and p= 0.013, respectively). Regarding the Notch signaling pathway, most ligands (JAG1 and DLL4) and receptors (NOTCH1 and NOTCH2) analyzed had increased expression in spheroids (p= 0.021, p= 0.028, p= 0.038 and p= 0.036, respectively). In Wnt pathway, we found higher expression levels of WNT3, CTNBB1 and GSK3B (p= 0.021, p= 0.008 and p= 0.021, respectively) in tumorspheres. No significant results were found for the rest of genes analyzed.
    
    Conclusion:
    Lung cancer spheroids from primary tumors and cell lines showed increased levels of genes related to CSCs properties. Genes belonging to Notch and Wnt signaling pathways were found to be more expressed in tumorspheres, suggesting that these pathways could be interesting lung-CSC targets. This work was supported in part, by grants RD12/0036/0025 from RTICC, and PI12-02838/PI15-0753 from ISCIII.