Identification and characterization of cells with cancer stem cell properties in human primary lung cancer cell lines

PLoS One. 2013;8(3):e57020. doi: 10.1371/journal.pone.0057020. Epub 2013 Mar 4.

Abstract

Lung cancer (LC) with its different subtypes is generally known as a therapy resistant cancer with the highest morbidity rate worldwide. Therapy resistance of a tumor is thought to be related to cancer stem cells (CSCs) within the tumors. There have been indications that the lung cancer is propagated and maintained by a small population of CSCs. To study this question we established a panel of 15 primary lung cancer cell lines (PLCCLs) from 20 fresh primary tumors using a robust serum-free culture system. We subsequently focused on identification of lung CSCs by studying these cell lines derived from 4 representative lung cancer subtypes such as small cell lung cancer (SCLC), large cell carcinoma (LCC), squamous cell carcinoma (SCC) and adenocarcinoma (AC). We identified a small population of cells strongly positive for CD44 (CD44(high)) and a main population which was either weakly positive or negative for CD44 (CD44(low/-)). Co-expression of CD90 further narrowed down the putative stem cell population in PLCCLs from SCLC and LCC as spheroid-forming cells were mainly found within the CD44(high)CD90(+) sub-population. Moreover, these CD44(high)CD90(+) cells revealed mesenchymal morphology, increased expression of mesenchymal markers N-Cadherin and Vimentin, increased mRNA levels of the embryonic stem cell related genes Nanog and Oct4 and increased resistance to irradiation compared to other sub-populations studied, suggesting the CD44(high)CD90(+) population a good candidate for the lung CSCs. Both CD44(high)CD90(+) and CD44(high)CD90(-) cells in the PLCCL derived from SCC formed spheroids, whereas the CD44(low/-) cells were lacking this potential. These results indicate that CD44(high)CD90(+) sub-population may represent CSCs in SCLC and LCC, whereas in SCC lung cancer subtype, CSC potentials were found within the CD44(high) sub-population.

Publication types

  • Research Support, Non-U.S. Gov't
  • Retracted Publication

MeSH terms

  • Adenocarcinoma / genetics*
  • Adenocarcinoma / metabolism
  • Adenocarcinoma / pathology
  • Adenocarcinoma of Lung
  • Animals
  • Antigens, CD / genetics
  • Antigens, CD / metabolism
  • Biomarkers, Tumor / genetics*
  • Biomarkers, Tumor / metabolism
  • Cadherins / genetics
  • Cadherins / metabolism
  • Carcinoma, Large Cell / genetics*
  • Carcinoma, Large Cell / metabolism
  • Carcinoma, Large Cell / pathology
  • Carcinoma, Squamous Cell / genetics*
  • Carcinoma, Squamous Cell / metabolism
  • Carcinoma, Squamous Cell / pathology
  • Gene Expression Regulation, Neoplastic*
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • Humans
  • Hyaluronan Receptors / genetics
  • Hyaluronan Receptors / metabolism
  • Immunophenotyping
  • Lung Neoplasms / genetics*
  • Lung Neoplasms / metabolism
  • Lung Neoplasms / pathology
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Nanog Homeobox Protein
  • Neoplastic Stem Cells / metabolism*
  • Neoplastic Stem Cells / pathology
  • Octamer Transcription Factor-3 / genetics
  • Octamer Transcription Factor-3 / metabolism
  • Primary Cell Culture
  • Small Cell Lung Carcinoma / genetics*
  • Small Cell Lung Carcinoma / metabolism
  • Small Cell Lung Carcinoma / pathology
  • Thy-1 Antigens / genetics
  • Thy-1 Antigens / metabolism
  • Vimentin / genetics
  • Vimentin / metabolism

Substances

  • Antigens, CD
  • Biomarkers, Tumor
  • CDH2 protein, human
  • Cadherins
  • Homeodomain Proteins
  • Hyaluronan Receptors
  • NANOG protein, human
  • Nanog Homeobox Protein
  • Octamer Transcription Factor-3
  • POU5F1 protein, human
  • Thy-1 Antigens
  • Vimentin

Grant support

This work was supported by a grant from the Norwegian research council (SFI-CAST). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.