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Correction: Generation and Characterisation of Cisplatin-Resistant Non-Small Cell Lung Cancer Cell Lines Displaying a Stem-Like Signature

DOI:10.1371/journal.pone.0233739
Authors:
Martin P. Barr at St James's Hospital & Trinity College Dublin
Martin P. Barr
  • St James's Hospital & Trinity College Dublin
Steven G Gray at St. James's Hospital
Steven G Gray
Andreas-Claudius Hoffmann at KMG Klinik Silbermühle GmbH
Andreas-Claudius Hoffmann
  • KMG Klinik Silbermühle GmbH
Ralf Axel Hilger at University Hospital Essen
Ralf Axel Hilger
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Abstract and Figures

[This corrects the article DOI: 10.1371/journal.pone.0054193.].
Figures - available via license: CC BY
Content may be subject to copyright.
Available via license: CC BY
Content may be subject to copyright.
CORRECTION
Correction: Generation and Characterisation
of Cisplatin-Resistant Non-Small Cell Lung
Cancer Cell Lines Displaying a Stem-Like
Signature
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
In Fig 11A of this article [1], the 12h MOR CisR image is duplicated in error as representing
the 4h MOR CisR result. In the revised Fig 11 provided here, the 4h MOR CisR has been
replaced with the correct data from the original experiment. The figure legend has been
updated to clarify methods used to obtain the quantitative results.
The original images underlying the updated Fig 11A are included in S1 and S2 Files of this
notice; S3–S6 Files include image and quantitative data supporting the results in Fig 11B. The
original data underlying other results in the article are available upon request from the
authors.
A member of PLOS ONE’s Editorial Board confirmed that the revised version of Fig 11 and
the data provided support the results as reported in the original article.
The authors apologize for the error in the published article.
PLOS ONE
PLOS ONE | https://doi.org/10.1371/journal.pone.0233739 May 21, 2020 1 / 3
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OPEN ACCESS
Citation: Barr MP, Gray SG, Hoffmann AC, Hilger
RA, Thomale J, O’Flaherty JD, et al. (2020)
Correction: Generation and Characterisation of
Cisplatin-Resistant Non-Small Cell Lung Cancer
Cell Lines Displaying a Stem-Like Signature. PLoS
ONE 15(5): e0233739. https://doi.org/10.1371/
journal.pone.0233739
Published: May 21, 2020
Copyright: ©2020 Barr et al. This is an open
access article distributed under the terms of the
Creative Commons Attribution License, which
permits unrestricted use, distribution, and
reproduction in any medium, provided the original
author and source are credited.
Fig 11. Cisplatin-DNA adduct formation and immunofluorescence. Lung cancer cell lines were treated with cisplatin for up to
24 h and fixed on Superfrost Gold Slides using ice-cold methanol. Cells were stained overnight at 4˚C using a primary antibody
that specifically recognizes CDDP-GpG DNA adducts (RC-18). Antibody binding was detected using an anti-rat Cy3
1
-labelled
antibody and counterstained using DAPI (1 μg/ml (w/v). Images were acquired on an Axioplan fluorescence microscope (A). The
quantitative data for adduct levels (B) in all cell lines (MOR, H460, A549 and SKMES-1) were measured using the ACAS-6.0 image
analysis system as described in the Materials & Methods section. This CCD camera-based methodology does not deliver
photographic images but directly measures the area of individual nuclei, their DNA content and the levels of Pt adduct-derived
fluorescence signals from automatically selected pixels. This is represented as relative numbers for all three parameters for 100–500
cells. These are then converted into AFU (arbitrary fluorescence units) values representing the relative adduct level of a single
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PLOS ONE | https://doi.org/10.1371/journal.pone.0233739 May 21, 2020 2 / 3
Supporting information
S1 File. Powerpoint file showing the original Fig 11A (with error), updated Fig 11A, and
the Cy3, DAPI, and DAPI Cy3 overlay images used in the MOR CisR (4h) panel of the
updated Fig 11A.
(PPT)
S2 File. MOR cells 4h, 12h and 24h (Cy3, DAPI and overlay).
(ZIP)
S3 File. H460 cells 4h, 12h and 24h (Cy3, DAPI and overlay).
(ZIP)
S4 File. A549 cells 4h, 12h and 24h (Cy3, DAPI and overlay).
(ZIP)
S5 File. SKMES-1 cells 4h, 12h and 24h (Cy3, DAPI and overlay).
(ZIP)
S6 File. Quantitative data (Adduct Kinetics).
(XLSX)
Reference
1. Barr MP, Gray SG, Hoffmann AC, Hilger RA, Thomale J, O’Flaherty JD, et al. (2013) Generation and
Characterisation of Cisplatin-Resistant Non-Small Cell Lung Cancer Cell Lines Displaying a Stem-Like
Signature. PLoS ONE 8(1): e54193. https://doi.org/10.1371/journal.pone.0054193 PMID: 23349823
nucleus. The graphics represented in Figure 11B show mean values of approximately 200 nuclei per time point ±95% confidence
intervals. In parallel with these analyses, cell images for all cell lines were acquired using Axioplan fluorescence microscopy.
https://doi.org/10.1371/journal.pone.0233739.g001
PLOS ONE
PLOS ONE | https://doi.org/10.1371/journal.pone.0233739 May 21, 2020 3 / 3
... Coexistence of CD44v3 and ALDH1A1 in head and neck cancer cells provides escape from apoptosis, promotes survival and proliferation through activation of downstream effectors such as Sox2, Nanog, and Oct4 [77]. Not only resistance to therapeutics but recent therapies such as ionizing radiation therapy or cisplatin may even cause CSC characteristics of cancer cells is another challenging point [78,79]. Pützer et al. discussed that rather than unilateral anti-CSC approaches strengthening patient's immune defense and heading toward individualized therapies CSC treatment can be successful [80]. ...
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Advanced Nanoparticle-Based Drug Delivery Systems and Their Cellular Evaluation for Non-Small Cell Lung Cancer Treatment
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Lung cancers, the number one cancer killer, can be broadly divided into small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC), with NSCLC being the most commonly diagnosed type. Anticancer agents for NSCLC suffer from various limitations that can be partly overcome by the application of nanomedicines. Nanoparticles is a branch within nanomedicine that can improve the delivery of anticancer drugs, whilst ensuring the stability and sufficient bioavailability following administration. There are many publications available in the literature exploring different types of nanoparticles from different materials. The effectiveness of a treatment option needs to be validated in suitable in vitro and/or in vivo models. This includes the developed nanoparticles, to prove their safety and efficacy. Many researchers have turned towards in vitro models that use normal cells or specific cells from diseased tissues. However, in cellular works, the physiological dynamics that is available in the body could not be mimicked entirely, and hence, there is still possible development of false positive or false negative results from the in vitro models. This article provides an overview of NSCLC, the different nanoparticles available to date, and in vitro evaluation of the nanoparticles. Different types of cells suitable for in vitro study and the important precautions to limit the development of false results are also extensively discussed.
Attenuation of lung cancer stem cell tumorigenesis and metastasis by cisplatin
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Purpose: To investigate the effect of cisplatin on the growth and metastasis abilities of lung cancer stem cells (CSCs) via molecular imaging. Materials and methods: The expression changes of lung CSCs cell marker in A549-Luc-C8 human non-small-cell lung cancer (NSCLC) cell line with or without cisplatin treatment were detected by flow cytometry. The tumorigenesis and metastasis abilities of A549-Luc-C8 cells were monitored both in vitro and in vivo, and the mechanism was assessed by gene sequencing. Results: About 1%-2% of CSCs were detected in A549-Luc-C8 cells and decreased CSCs percentage was observed after cisplatin treatment. Attenuated tumorigenesis and metastasis abilities of A549-Luc-C8 cells were found in cisplatin treated group. Conclusions: Decreased percentage of CSCs in A549-Luc-C8 cells can be induced by cisplatin treatment, which may partly be attributed to the attenuated expression of growth factors.
CD10 as a novel marker of therapeutic resistance and cancer stem cells in head and neck squamous cell carcinoma
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Full-text available
  • May 2014
Background: Cancer stem cells (CSCs) are responsible for treatment failure. However, their identification and roles in resistance are not well established in head and neck squamous cell carcinoma (HNSCC). Methods: Three HNSCC cell lines (FaDu, Detroit562 and BICR6) were treated with cisplatin or radiation. Cell surface antigens were analysed by LyoPlate, a novel cell surface antigen array. The expression levels of antigens highly expressed after treatments were further compared between cisplatin-resistant Detroit562 cells and its parental line. Association of the candidate antigen with CSCs properties, namely sphere formation and in vivo tumourigenicity, was also examined. Results: CD10, CD15s, CD146 and CD282 were upregulated across the treated cell lines, while the increased expression of CD10 was prominent in the cisplatin-resistant cell line. Isolation mediated by FACS revealed that the CD10-positive subpopulation was more refractory to cisplatin, fluorouracil and radiation than the CD10-negative subpopulation. It also showed an increased ability to form spheres in vitro and tumours in vivo. Moreover, the CD10-positive subpopulation expressed the CSC marker OCT3/4 at a higher level than that in the CD10-negative subpopulation. Conclusions: CD10 is associated with therapeutic resistance and CSC-like properties of HNSCC. CD10 may serve as a target molecule in the treatment of refractory HNSCC.
Generation and Characterisation of Cisplatin-Resistant Non-Small Cell Lung Cancer Cell Lines Displaying a Stem-Like Signature
Article
Full-text available
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. 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. 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. 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.
The graphics represented in Figure 11B show mean values of approximately 200 nuclei per time point ±95% confidence intervals. In parallel with these analyses, cell images for all cell lines were acquired using Axioplan fluorescence microscopy
nucleus. The graphics represented in Figure 11B show mean values of approximately 200 nuclei per time point ±95% confidence intervals. In parallel with these analyses, cell images for all cell lines were acquired using Axioplan fluorescence microscopy. https://doi.org/10.1371/journal.pone.0233739.g001