Generation and characterisation of Cisplatin-resistant non-small cell lung cancer cell lines displaying a stem-like signature.
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Journal ArticleDate:
2013Access:
openAccessCitation:
Barr MP, Gray SG, Hoffmann AC, Hilger RA, Thomale J, O Flaherty JD, Fennell DA, Richard D, O Leary JJ, O'Byrne KJ., Generation and characterisation of Cisplatin-resistant non-small cell lung cancer cell lines displaying a stem-like signature., PloS one, 8, 1, 2013, e54193Download Item:
Abstract:
Introduction:
Inherent and acquired cisplatin resistance reduces the effectiveness of this agent in the management of non-
small cell lung cancer (NSCLC). Understanding the molecular mechanisms underlying this process may result in the
development of novel agents to enhance the sensitivity of cisplatin.
Methods:
An isogenic model of cisplatin resistance was generated in a panel of NSCLC cell lines (A549, SKMES-1, MOR,
H460). Over a period of twelve months, cisplatin resistant (CisR) cell lines were derived from original, age-matched parent
cells (PT) and subsequently characterized. Proliferation (MTT) and clonogenic survival assays (crystal violet) were carried out
between PT and CisR cells. Cellular response to cisplatin-induced apoptosis and cell cycle distribution were examined by
FACS analysis. A panel of cancer stem cell and pluripotent markers was examined in addition to the EMT proteins, c-Met and
b
-catenin. Cisplatin-DNA adduct formation, DNA damage (
c
H2AX) and cellular platinum uptake (ICP-MS) was also assessed.
Results:
Characterisation studies demonstrated a decreased proliferative capacity of lung tumour cells in response to
cisplatin, increased resistance to cisplatin-induced cell death, accumulation of resistant cells in the G0/G1 phase of the cell
cycle and enhanced clonogenic survival ability. Moreover, resistant cells displayed a putative stem-like signature with
increased expression of CD133
+
/CD44
+
cells and increased ALDH activity relative to their corresponding parental cells. The
stem cell markers, Nanog, Oct-4 and SOX-2, were significantly upregulated as were the EMT markers, c-Met and
b
-catenin.
While resistant sublines demonstrated decreased uptake of cisplatin in response to treatment, reduced cisplatin-GpG DNA
adduct formation and significantly decreased
c
H2AX foci were observed compared to parental cell lines.
Conclusion:
Our results identified cisplatin resistant subpopulations of NSCLC cells with a putative stem-like signature,
providing a further understanding of the cellular events associated with the cisplatin resistance phenotype in lung cancer.
Author's Homepage:
http://people.tcd.ie/olearyjjhttp://people.tcd.ie/barrma
http://people.tcd.ie/grayst
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PUBLISHEDType of material:
Journal ArticleSeries/Report no:
PloS one8
1
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Full text availableKeywords:
cisplatin resistanceSubject (TCD):
Cancer , Biomedical sciencesDOI:
http://dx.doi.org/ 10.1371/journal.pone.0054193Metadata
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