Induction and repair of DNA double-strand breaks using constant-field gel electrophoresis and apoptosis as predictive markers for sensitivity of cancer cells to cisplatin

Saleh, Ekram; El‐Awady, Raafat; Anis, Noha; El-Sharkawy, Nahla

doi:10.1016/j.biopha.2012.07.001

Cited by 9 publications

(7 citation statements)

References 34 publications

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“…To overcome the shortcomings of using mutational status and genomic instability to predict platinum response, we and other groups have developed phenotypic assays to predict response after treatment. For example, Saleh et al treated cancer cell lines with cisplatin and quantitated the level of double strand breaks via constant-field gel electrophoresis and found a correlation between double strand breaks and cisplatin sensitivity . In the current study, we treated breast cancer cell lines with carboplatin or oxaliplatin and quantitated the formation of drug-DNA adducts via accelerator mass spectrometry (AMS) and found correlation with platinum sensitivity.…”

Section: Introductionmentioning

confidence: 70%

Correlation of Platinum Cytotoxicity to Drug-DNA Adduct Levels in a Breast Cancer Cell Line Panel

Wang

Scharadin

Zimmermann

et al. 2018

Chem. Res. Toxicol.

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Platinum drugs, including carboplatin and oxaliplatin, are commonly used chemotherapy drugs that kill cancer cells by forming toxic drug-DNA adducts. These drugs have a proven, but modest, efficacy against several aggressive subtypes of breast cancer but also cause several side effects that can lead to the cessation of treatment. There is a clinical need to identify patients who will respond to platinum drugs in order to better inform clinical decision making. Diagnostic microdosing involves dosing patients or patient samples with subtherapeutic doses of radiolabeled platinum followed by measurement of platinum-DNA adducts in blood or tumor tissue and may be used to predict patient response. We exposed a panel of six breast cancer cell lines to 14C-labeled carboplatin or oxaliplatin at therapeutic and microdose (1% therapeutic dose) concentrations for a range of exposure lengths and isolated DNA from the cells. The DNA was converted to graphite, and measurement of radiocarbon due to platinum-DNA adduction was quantified via accelerator mass spectrometry (AMS). We observed a linear correlation in adduct levels between the microdose and therapeutic dose, and the level of platinum-DNA adducts corresponded to cell line drug sensitivity for both carboplatin and oxaliplatin. These results showed a clear separation in adduct levels between the sensitive and resistant groups of cell lines that could not be fully explained or predicted by changes in DNA repair rates or mutations in DNA repair genes. Further, we were able to quantitate oxaliplatin-DNA adducts in the blood and tumor tissue of a metastatic breast cancer patient. Together, these data support the use of diagnostic microdosing for predicting patient sensitivity to platinum. Future studies will be aimed at replicating this data in a clinical feasibility trial.

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Section: Introductionmentioning

confidence: 70%

Correlation of Platinum Cytotoxicity to Drug-DNA Adduct Levels in a Breast Cancer Cell Line Panel

Wang

Scharadin

Zimmermann

et al. 2018

Chem. Res. Toxicol.

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“…The mode of action of cisplatin is primarily through the formation of DNA adducts and double strand breaks that result in the activation of signaling pathways that ultimately lead to apoptosis. [35][36][37] In previous work, glycolysis in MCF7 cells has been shown to increase following treatment with cisplatin 38 and we therefore investigated whether treatment with cisplatin could lead to a change in the NADH°uorescence lifetime parameters. Our preliminary results presented here in Sec.…”

Section: Discussionmentioning

confidence: 99%

An automated multiwell plate reading flim microscope for live cell autofluorescence lifetime assays

Kelly

Warren

Kumar

et al. 2014

J. Innov. Opt. Health Sci.

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Fluorescence lifetime imaging (FLIM) is increasingly used to read out cellular auto°uorescence originating from the coenzyme NADH in the context of investigating cell metabolic state. We present here an automated multiwell plate reading FLIM microscope optimized for UV illumination with the goal of extending high content°uorescence lifetime assays to readouts of metabolism. We demonstrate its application to automated cellular auto°uorescence lifetime imaging and discuss the key practical issues associated with its implementation. In particular, we illustrate its capability to read out the NADH-lifetime response of cells to metabolic modulators, thereby illustrating the potential of the instrument for cytotoxicity studies, assays for drug discovery and strati¯ed medicine.

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“…In addition, the effect of CDDP-loaded hydrogels on the cell cycle of MCF-7 cells was investigated by detecting DNA content after staining with PI. As illustrated in Figure 4B, CDDP-loaded hydrogels induced predominant S phase arrest of MCF-7 cells (50.10%) as compared with the PBS group (38.35%) [32][33][34], while the percentages of MCF-7 cells incubated with blank hydrogels in various phases were comparable to those of the PBS group. This further manifested the cytocompatibility of the iEDDA-based polypeptide hydrogels.…”

Section: In Vitro Tumor Cell Inhibition By Cddp-loaded Hydrogelsmentioning

confidence: 82%

Injectable Click Polypeptide Hydrogels via Tetrazine-Norbornene Chemistry for Localized Cisplatin Release

Zhang

Chen

2020

Polymers

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Injectable, covalently cross-linked hydrogels have been widely investigated in drug delivery systems due to their superior mechanical properties and long-term stability. Conventional covalently cross-linked hydrogels are formed by chemical reactions that may interfere with natural biochemical processes. In this work, we developed an injectable polypeptide hydrogel via an inverse electron demand Diels-Alder reaction between norbornene modified poly(L-glutamic acid) (PLG-Norb) and tetrazine functionalized four-arm poly(ethylene glycol) (4aPEG-T) for localized release of cisplatin (CDDP). The rapid and bioorthogonal click reaction allowed for hydrogel formation within a few minutes after mixing the two polymer solutions in phosphate buffer saline (PBS). Dynamic mechanical analysis suggested that the storage modulus of the hydrogel could be readily tuned by changing the polymer concentration and the molar ratio of the two functional groups. The carboxyl groups of PLG-Norb were used to form polymer–metal complexation with CDDP, and the controlled release of the antitumor drug was achieved in PBS. The CDDP-loaded hydrogel displayed an antitumor effect against MCF-7 cells in vitro, through S phase cell cycle arrest. After subcutaneous injection in rats, the hydrogel was rapidly formed in situ and showed good stability in vivo. In an MCF-7-bearing nude mice model, the CDDP-loaded hydrogel exhibited an improved antitumor effect with reduced systemic toxicity. Overall, the injectable click polypeptide hydrogel shows considerable potential as a platform for localized and sustained delivery of antitumor drugs.

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Induction and repair of DNA double-strand breaks using constant-field gel electrophoresis and apoptosis as predictive markers for sensitivity of cancer cells to cisplatin

Cited by 9 publications

References 34 publications

Correlation of Platinum Cytotoxicity to Drug-DNA Adduct Levels in a Breast Cancer Cell Line Panel

Correlation of Platinum Cytotoxicity to Drug-DNA Adduct Levels in a Breast Cancer Cell Line Panel

An automated multiwell plate reading flim microscope for live cell autofluorescence lifetime assays

Injectable Click Polypeptide Hydrogels via Tetrazine-Norbornene Chemistry for Localized Cisplatin Release

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