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Generation and characterisation of Cisplatin-resistant non-small cell lung cancer cell lines displaying a stem-like signature.
BARR, MARTIN; O'LEARY, JOHN; GRAY, STEVEN
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.
Keyword(s): cisplatin resistance; Cancer; Biomedical sciences
Publication Date:
2013
Type: Journal article
Peer-Reviewed: Yes
Language(s): English
Institution: Trinity College Dublin
Citation(s): 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, e54193
First Indexed: 2015-03-07 05:50:22 Last Updated: 2015-05-02 05:19:45