Resazurin assay of radiation response in cultured cells

Anoopkumar‐Dukie, Shailendra; Carey, John J.; Conere, T. J.; O'Sullivan, E.; Pelt, F N van; Allshire, Ashley

doi:10.1259/bjr/54004230

Cited by 152 publications

(103 citation statements)

References 8 publications

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“…previously shown to yield comparable results with the clonogenic assay [11]. Irradiation did not affect cell capacity to reduce the dye and differences reflected cell number only (not shown).…”

Section: Resazurin (Alamar Blue) Assaysupporting

confidence: 66%

“…This method yields results comparable to both the standard clonogenic assay and the DNA synthesis rate as measured by BrdU incorporation [11], as well as to DNA content (measured by the CyQUANT TM assay; unpublished). Irradiation of all three cell lines at 6.2 Gy had limited impact under hypoxia while irradiation under ambient oxygen markedly impaired cell proliferation, illustrating significant oxygen-dependent radiosensitivity ( Figure 1).…”

Section: Ptp Involvement In Oxygen-dependent Irradiation Injurymentioning

confidence: 83%

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Mitochondrial modulation of oxygen-dependent radiosensitivity in some human tumour cell lines

Anoopkumar‐Dukie

Conere²,

Sisk³

et al. 2009

BJR

Self Cite

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ABSTRACT. Oxygen-dependent radiosensitivity of tumour cells reflects direct oxidative damage to DNA, but non-nuclear mechanisms including signalling pathways may also contribute. Mitochondria are likely candidates because not only do they integrate signals from each of the main kinase pathways but mitochondrial kinases responsive to oxidative stress communicate to the rest of the cell. Using pharmacological and immunochemical methods, we tested the role of mitochondrial permeability transition (MPT) and the Bcl-2 proteins in oxygen-dependent radiosensitivity. Drug-treated or untreated cervical cancer HeLa, breast cancer MCF-7 and melanoma MeWo cell lines were irradiated at 6.2 Gy under normoxic and hypoxic conditions then allowed to proliferate for 7 days. The MPT blocker cyclosporin A (2 mM) strongly protected HeLa but not the other two lines against oxygen-dependent radiosensitivity. By contrast, bongkrekic acid (50 mM), which blocks MPT by targeting the adenine nucleotide transporter, had only marginal effect and calcineurin inhibitor FK-506 (0.1 mM) had none. Nor was evidence found for the modulation of oxygen-dependent radiosensitivity by Bax/Bcl-2 signalling, mitochondrial ATP-dependent potassium (mitoK ATP ) channels or mitochondrial Ca 2+ uptake. In conclusion, calcineurinindependent protection by cyclosporin A suggests that MPT but not mitoK ATP or the mitochondrial apoptosis pathway plays a causal role in oxygen-dependent radiosensitivity of HeLa cells. Targeting MPT may therefore improve the effectiveness of radiotherapy in some solid tumours.

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Section: Resazurin (Alamar Blue) Assaysupporting

confidence: 66%

Section: Ptp Involvement In Oxygen-dependent Irradiation Injurymentioning

confidence: 83%

Mitochondrial modulation of oxygen-dependent radiosensitivity in some human tumour cell lines

Anoopkumar‐Dukie

Conere²,

Sisk³

et al. 2009

BJR

Self Cite

View full text Add to dashboard Cite

show abstract

“…The Alamar Blue assay was chosen as it allows repeated measure-ment of cell proliferation without destruction of the cultures, and it gives proliferation measurements equivalent to those carried out with BrdU incorporation [37,38]. In neonatal SC the number of passages does not appear to negatively influence the growth rate of the cells.…”

Section: Discussionmentioning

confidence: 99%

Morphological, molecular and functional differences of adult bone marrow- and adipose-derived stem cells isolated from rats of different ages

Mantovani

Raimondo

Haneef

et al. 2012

Experimental Cell Research

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“…analyze the toxicity of compounds in cell culture [11]. The analysis is based on the ability of metabolically active cells to restore resazurin (a blue product) to resorufin (a pink color product, a fluorophore with absorption and emission maxima at 571 and 586 nm, respectively).…”

Section: Test For Vitality With Resazurin Resazurin Is An Oxidation-mentioning

confidence: 99%

Master-regulators driving resistance of non-small cell lung cancer cells to p53 reactivator Nutlin-3

Boyarskikh

Pintus²,

Mandrik³

et al. 2017

Virtual Biology

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SummaryThe tumor suppressor protein p53 is one of the most important targets in various cancers including non-small cell lung cancer (NSCLC). The small molecule Nutlin-3 reactivates p53 in cancer cells by interacting with the complex between p53 and its repressor Mdm-2, which leads to increased apoptosis of cancer cells. Nevertheless, clinical and experimental studies of Nutlin-3 have shown that in some cases cancer cells are not sensitive to this compound. In this paper we studied possible mechanisms of such a resistance of cancer cells to Nutlin-3 through increased activity of specific pro-survival pathways that appeared to be more active in the resistant cells, as compared to the cells sensitive to the treatment by this compound. Using genome-wide gene expression profiling we compared several NSCLC cell lines. Nutlin-3 resistant cell lines showed a significantly different gene expression in comparison with sensitive cell lines before and after treatment by Nutlin-3.Using original bioinformatics approaches we analyzed the revealed gene expression patterns and identified complexes of transcription factors specifically regulating different expression of genes in the resistant cell lines. Analysis of the signal transduction network upstream from transcription factors allowed us to identify potential master-regulators responsible for 2 maintaining this resistant state. Among them the most promising was mTOR acting in the context of the activated PI3K pathway. With the help of chemical inhibitors, we were able to validate these findings experimentally. The Nutlin-3 resistant cell lines showed the highest sensitivity to the inhibitor of mTOR with PI3K, which lead to the rapid death of these cells.

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Resazurin assay of radiation response in cultured cells

Cited by 152 publications

References 8 publications

Mitochondrial modulation of oxygen-dependent radiosensitivity in some human tumour cell lines

Mitochondrial modulation of oxygen-dependent radiosensitivity in some human tumour cell lines

Morphological, molecular and functional differences of adult bone marrow- and adipose-derived stem cells isolated from rats of different ages

Master-regulators driving resistance of non-small cell lung cancer cells to p53 reactivator Nutlin-3

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