62P - Novel therapeutic strategies in NSCLC: Lung-tumorspheres as an effective in vitro screening platform

Date 15 April 2016
Event European Lung Cancer Conference 2016 (ELCC) 2016
Session Poster lunch
Topics Thoracic malignancies
Translational Research
Basic Principles in the Management and Treatment (of cancer)
Presenter Ester Munera Maravilla
Citation Journal of Thoracic Oncology (2016) 11 (supplement 4): S57-S166. S1556-0864(16)X0004-4
Authors E. Munera Maravilla1, A. Herreros Pomares1, S. Calabuig-Fariñas2, E. Jantus Lewintre3, R. Lucas4, R. Guijarro5, A.I. Martinez Madriaga5, A. Blasco Cordellat6, C. Camps7
  • 1Laboratorio De Oncologia Molecular, Fundación para la Investigación del Hospital General de Valencia, 46014 - Valencia/ES
  • 2Department Of Pathology, Universitat De València, Laboratorio de Oncologia Molecular, Fundación para la Investigación del Hospital General Universitario de Valencia, 46014 - Valencia/ES
  • 3Department Of Biotechnology, Universidad Politécnica De Valencia, Molecular Oncology Laboratory, General University Hospital Research Foundation, Valencia/ES
  • 4Departamento De Historia De La Ciencia, Universidad de Valencia, Valencia/ES
  • 5Department Of Thoracic Surgery, Hospital General Universitario Valencia, Valencia/ES
  • 6Servicio De Oncologia Medica, Hospital General Universitario Valencia, 46018 - Valencia/ES
  • 7Department 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

Resistance to treatment is one of the causes influencing the high mortality of lung cancer. This feature seems to be linked to a subpopulation known as Cancer Stem Cells (CSCs), which are able to grow as spheroids (suspension culture). The aim of the study was to obtain tumorspheres from lung cancer cell lines and primary tumor samples from non-small cell lung cancer (NSCLC) patients and to use them as an in vitro platform for drug screening.

Methods

Cells from resected NSCLC tumor samples and lung cancer cell lines were grown in monolayer and as spheroids. Cultured cells were used: (i) to compare the cytotoxic effect of anticancer drugs in adherent vs lung-tumorspheres and (ii) to perform a high-throughput screening with commercial chemical libraries. Briefly, cells were plated at the desired density in 200 µl of medium in 96-well plates and compounds were added at 4 different concentrations (n = 3) Cell viability was measured after 72 h, using MTS Assay. Cell viability was normalized to the respective mock-treated control cells and presented as percent of control.

Results

Cells cultured in serum-free conditions were able to form spheroids, such as stem-like cells. Under these culture conditions, classical anticancer drugs (cisplatin, paclitaxel, doxorubicin) exhibited mild or null cytotoxic effects on H1650, H1993, A549, PC09 spheroids. The same behavior was observed in lungspheres derived from NSCLC primary tumors at concentrations ranging from 1 to 10 µM. Moreover, we performed a high-throughput screening with commercial chemical libraries and remarkably, only three compounds reduced the number of viable tumorspheres. Characterization of these compounds is ongoing.

Conclusions

Our data suggest that lung-tumorspheres showed resistance to classical anticancer drugs, strengthening its possible use as a short-term culture platform for a simple, and cost-effective screening to investigate novel therapeutic approaches. In this setting, three compounds were identified as promising therapeutic agents on lung tumorspheres, but confirmatory data are still necessary.

Clinical trial identification

Legal entity responsible for the study

Fundación para la Investigación del Hospital General Universitario de Valencia

Funding

Supported in part, by a grant [RD12/0036/0025] from RTICC, PI12–02838 from ISCIII and SEOM/2012.

Disclosure

All authors have declared no conflicts of interest.