Cell death stages in single apoptotic and necrotic cells monitored by Raman microspectroscopy

Brauchle, Eva; Thude, Sibylle; Brucker, Sara Y.; Schenke‐Layland, Katja

doi:10.1038/srep04698

Cited by 246 publications

(203 citation statements)

References 47 publications

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“…17 are secondary necrotic cells. 18 Our results showed that there has been less necrosis in our study compared to the results of Miura and Shinohara. 17 The inhibition of A549 cells has been induced by silver nitrate.…”

Section: Discussioncontrasting

confidence: 52%

The apoptotic and genomic studies on A549 cell line induced by silver nitrate

2017

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Lung cancer is the leading cause of male cancer deaths worldwide. Metal-based anticancer drugs have evolved significantly during the past decades. Recently, silver ions have been investigated for their anticancer effects. We aimed to study the time-course cytotoxic effects of silver nitrate on A549 adenocarcinomic human alveolar basal epithelial cells to provide insights into the molecular-level understanding of growth suppression mechanism involved in apoptosis. The influences of silver nitrate were studied via MTT assay, flow cytometry, immunocytochemical, confocal and transmission electron microscopy, and microarray assays. Silver nitrate showed inhibitory effects against A549 cells in a dose-and time-dependent manner for 24, 48, and 72 h and induced apoptosis. The early and late apoptotic cells and depolarized mitochondrial membrane potential were determined by the half-maximal inhibitory concentration (IC 50 ) value of silver nitrate treated for 72 h. But cysteinyl aspartate proteinase-3 was not activated for 72 h. Furthermore, IC 50 value of silver nitrate also induced apoptosis according to immunocytochemical assays for 72 h. The downregulated CCNY, HNRNPL, ASF1B, PIAS4, HNRNPH1, EIF2C2, TAF15, FOXC1, LEP, and PCB2 genes administered with silver nitrate IC 50 were identified as apoptosis-leading genes. Silver nitrate may be a suitable therapeutic agent against lung cancer.

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Section: Discussioncontrasting

confidence: 52%

The apoptotic and genomic studies on A549 cell line induced by silver nitrate

2017

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“…Note that the feature at 1425 cm −1 is increased because of the contribution of the DOX peak at the same wavenumber but after DOX subtraction it can be clearly seen that this peak is diminished (see These results are in accordance with the expected changes in cell nuclei, related to the DOX mechanism of action, by which DNA synthesis is blocked by intercalation causing changes in DNA conformation (decrease of DNA B form) 13 inducing early cell apoptosis. 51,52 (Full details of cellular peak assignments are given in Table 1). A shift towards lower wavenumbers is observed for the peak at 1095 cm −1 , corresponding to O-P-O stretching, (shown in the expanded section of Fig.…”

Section: Raman Micro Spectroscopymentioning

confidence: 99%

Raman micro spectroscopy for in vitro drug screening: subcellular localisation and interactions of doxorubicin

Farhane

Bonnier

Casey

et al. 2015

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Raman spectroscopy is used for the localization and tracking of chemotherapeutic drug, doxorubicin, in the intracellular environment of lung cancer cell line. Results show the potential of the technique to monitor the mechanisms of action and response on a molecular level, with subcellular resolution.Please check this proof carefully. Our staff will not read it in detail after you have returned it.Translation errors between word-processor files and typesetting systems can occur so the whole proof needs to be read. Please pay particular attention to: tabulated material; equations; numerical data; figures and graphics; and references. If you have not already indicated the corresponding author(s) please mark their name(s) with an asterisk. Please e-mail a list of corrections or the PDF with electronic notes attached -do not change the text within the PDF file or send a revised manuscript. Corrections at this stage should be minor and not involve extensive changes. All corrections must be sent at the same time.Please bear in mind that minor layout improvements, e.g. in line breaking, table widths and graphic placement, are routinely applied to the final version.We will publish articles on the web as soon as possible after receiving your corrections; no late corrections will be made. Please ensure that all queries are answered when returning your proof corrections so that publication of your article is not delayed. Vibrational spectroscopy, including Raman spectroscopy, has been widely used over the last few years to explore potential biomedical applications. Indeed, Raman spectroscopy has been demonstrated to be a powerful non-invasive tool in cancer diagnosis and monitoring. In confocal microscopic mode, the technique is also a molecularly specific analytical tool with optical resolution which has potential applications in subcellular analysis of biochemical processes, and therefore as an in vitro screening tool of the efficacy and mode of action of, for example, chemotherapeutic agents. In order to demonstrate and explore the potential in this field, established, model chemotherapeutic agents can be valuable. In study paper, Raman spectroscopy coupled with confocal microscopy were used for the localization and tracking of the commercially available drug, doxorubicin (DOX), in the intracellular environment of the lung cancer cell line, A549. Cytotoxicity assays were employed to establish clinically relevant drug doses for 24 h exposure, and confocal laser scanning fluorescence microscopy was conducted in parallel with Raman spectroscopy profiling to confirm the drug internalisation and localisation. Multivariate statistical analysis, consisting of PCA ( principal components analysis) was used to highlight doxorubicin interaction with cancer cells and spectral variations due to its effects before and after DOX spectral features subtraction from nuclear and nucleolar spectra, were compared to non-exposed control spectra. Results show that Raman micro spectroscopy is not only able to detect doxorubicin inside cells an...

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“…36 It has been suggested before that the decrease of the region may correlate with the progress of internucleosomal DNA cleavage. 37,38 Together, our results indicate that the decreases in Raman intensities of activated mature DNA are related to apoptosis. These findings also suggest that Raman spectroscopy can be used as a label-free and noninvasive optical technique to determine the status of activated mature CD8þ T cells.…”

Section: Discussionmentioning

confidence: 61%

Investigation of biochemical property changes in activation-induced CD 8 + T cell apoptosis using Raman spectroscopy

et al. 2015

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Abstract. The study was to investigate the changes in biochemical properties of activated mature CD8þ T cells related to apoptosis at a molecular level. We confirmed the activation and apoptosis of CD8þ T cells by fluorescence-activated cell sorting and atomic force microscopy and then performed Raman spectral measurements on activated mature CD8þ T cells and cellular deoxyribose nucleic acid (DNA). In the activated mature CD8þ T cells, there were increases in protein spectra at 1002 and 1234 cm −1 . In particular, to assess the apoptosisrelated DNA spectral signatures, we investigated the spectra of the cellular DNA isolated from resting and activated mature CD8þ T cells. Raman spectra at 765 to 786 cm −1 and 1053 to 1087 cm −1 were decreased in activated mature DNA. In addition, we analyzed Raman spectrum using the multivariate statistical method including principal component analysis. Raman spectra of activated mature DNA are especially well-discriminated from those of resting DNA. Our findings regarding the biochemical and structural changes associated with apoptosis in activated mature T cells and cellular DNA according to Raman spectroscopy provide important insights into allospecific immune responses generated after organ transplantation, and may be useful for therapeutic manipulation of the immune response. © The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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Cell death stages in single apoptotic and necrotic cells monitored by Raman microspectroscopy

Cited by 246 publications

References 47 publications

The apoptotic and genomic studies on A549 cell line induced by silver nitrate

The apoptotic and genomic studies on A549 cell line induced by silver nitrate

Raman micro spectroscopy for in vitro drug screening: subcellular localisation and interactions of doxorubicin

Investigation of biochemical property changes in activation-induced CD 8 + T cell apoptosis using Raman spectroscopy

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