Generation and characterisation of cisplatin-resistant non-small cell lung cancer cell lines displaying a stem-like signature.
<h4>Introduction</h4>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 sen...
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2013
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oai:doaj.org-article:18af4cd6dbdb4d73aa78e1b2488d6a3e2021-11-18T08:01:02ZGeneration and characterisation of cisplatin-resistant non-small cell lung cancer cell lines displaying a stem-like signature.1932-620310.1371/journal.pone.0054193https://doaj.org/article/18af4cd6dbdb4d73aa78e1b2488d6a3e2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23349823/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Introduction</h4>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.<h4>Methods</h4>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 β-catenin. Cisplatin-DNA adduct formation, DNA damage (γH2AX) and cellular platinum uptake (ICP-MS) was also assessed.<h4>Results</h4>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 β-catenin. While resistant sublines demonstrated decreased uptake of cisplatin in response to treatment, reduced cisplatin-GpG DNA adduct formation and significantly decreased γH2AX foci were observed compared to parental cell lines.<h4>Conclusion</h4>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.Martin P BarrSteven G GrayAndreas C HoffmannRalf A HilgerJuergen ThomaleJohn D O'FlahertyDean A FennellDerek RichardJohn J O'LearyKenneth J O'ByrnePublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 1, p e54193 (2013) |
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Medicine R Science Q Martin P Barr Steven G Gray Andreas C Hoffmann Ralf A Hilger Juergen Thomale John D O'Flaherty Dean A Fennell Derek Richard John J O'Leary Kenneth J O'Byrne Generation and characterisation of cisplatin-resistant non-small cell lung cancer cell lines displaying a stem-like signature. |
| description |
<h4>Introduction</h4>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.<h4>Methods</h4>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 β-catenin. Cisplatin-DNA adduct formation, DNA damage (γH2AX) and cellular platinum uptake (ICP-MS) was also assessed.<h4>Results</h4>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 β-catenin. While resistant sublines demonstrated decreased uptake of cisplatin in response to treatment, reduced cisplatin-GpG DNA adduct formation and significantly decreased γH2AX foci were observed compared to parental cell lines.<h4>Conclusion</h4>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. |
| format |
article |
| author |
Martin P Barr Steven G Gray Andreas C Hoffmann Ralf A Hilger Juergen Thomale John D O'Flaherty Dean A Fennell Derek Richard John J O'Leary Kenneth J O'Byrne |
| author_facet |
Martin P Barr Steven G Gray Andreas C Hoffmann Ralf A Hilger Juergen Thomale John D O'Flaherty Dean A Fennell Derek Richard John J O'Leary Kenneth J O'Byrne |
| author_sort |
Martin P Barr |
| title |
Generation and characterisation of cisplatin-resistant non-small cell lung cancer cell lines displaying a stem-like signature. |
| title_short |
Generation and characterisation of cisplatin-resistant non-small cell lung cancer cell lines displaying a stem-like signature. |
| title_full |
Generation and characterisation of cisplatin-resistant non-small cell lung cancer cell lines displaying a stem-like signature. |
| title_fullStr |
Generation and characterisation of cisplatin-resistant non-small cell lung cancer cell lines displaying a stem-like signature. |
| title_full_unstemmed |
Generation and characterisation of cisplatin-resistant non-small cell lung cancer cell lines displaying a stem-like signature. |
| title_sort |
generation and characterisation of cisplatin-resistant non-small cell lung cancer cell lines displaying a stem-like signature. |
| publisher |
Public Library of Science (PLoS) |
| publishDate |
2013 |
| url |
https://doaj.org/article/18af4cd6dbdb4d73aa78e1b2488d6a3e |
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