Quantitative evaluation of cellular uptake, DNA incorporation and adduct formation in cisplatin sensitive and resistant cell lines: Comparison of different Pt-containing drugs

Corte‐Rodríguez, Mario; Espina, Marta; Sierra, María R.; Blanco, Elisa; Ames, Tyler D.; Montes-Bayón, Marı́a; Sanz‐Medel, Alfredo

doi:10.1016/j.bcp.2015.08.112

Cited by 56 publications

(57 citation statements)

References 27 publications

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“…10,22 Despite similarities in cell viability with the GM04312 cells, the sensitivity of the A2780 cells to cisplatin cannot be linked to a deficiency of DNA repair. However, it might be related to their higher Pt influx, as described, 27,37,42,43 even for other platinum chemicals, 44 which could be related to high levels of the copper transport protein CTR1, 23 although recently the influence of this type of protein was considered controversial. 45 Nevertheless, since high Pt influx was not accompanied by high levels of induced DNA adducts, the drug effects on cell viability in these A2780 cells must be related to cisplatin activity in the cell cytoplasm, and not specifically related to apoptosis, because the apoptotic cell number did not significantly change at different Pt concentrations.…”

Section: Discussionmentioning

confidence: 99%

“…The cell pellets were weighed and were digested with 500 mL of 65% HNO 3 (Suprapur, Merck) for 1 hour at 70 1C, and then with 500 mL of 30% H 2 O 2 for 3 h at 70 1C, to produce transparent solutions suitable for Pt determination by ICP-MS. 27 The Pt concentration obtained thereafter was corrected by the dry cell mass. Two independent experiments were performed for each cell line.…”

Section: Total Intracellular Pt Quantitationmentioning

confidence: 99%

“…The 31 P signal was calibrated using digested calf thymus DNA standards, as previously described. 27 DNA adduct identification and quantitation were performed by HPLC-ICP-MS. 24 Briefly, DNA from each extraction was incubated with 5600U of S1-Nuclease overnight. The digestion products were passed through a 10 kDa pore-size ultrafiltration membrane (Amicon Ultra-Millipore) at 14 000g for 15 min to remove the enzyme.…”

Section: Cisplatin-induced Dna Adduct Quantitationmentioning

confidence: 99%

“…The gradient was set as follows: 0-5 min 3% B, 5-35 min 15% B, and 35-40 min 15% B. Quantitation was performed by ICP-MS detection (Pt monitoring). 27 At least three independent experiments were performed for each AT time and cell line.…”

Section: Cisplatin-induced Dna Adduct Quantitationmentioning

confidence: 99%

“…26 Moreover, we have recently developed an approach to quantitatively compare intracellular platinum content and adducted DNA among cell lines. 27 Considering all this, in this work we have tried to evaluate the cross-correlation of cisplatin-induced DNA adducts, intracellular Pt-content, cell cycle progression and drug-induced genomic instability (measured with the Comet assay), using four human cultured cell lines, A549, GM04312, A2780 and A2780cis, with different origins and cisplatin sensitivities. 23,28,29 Our final aim was to address the most accurate strategy to predict cisplatin sensitivity/resistance after short-time treatments with doses around the peak plasma concentration achieved in Pt-treated cancer patients.…”

Section: Introductionmentioning

confidence: 99%

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Cisplatin resistance in cell models: evaluation of metallomic and biological predictive biomarkers to address early therapy failure

et al. 2017

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Cisplatin, one of the most extensively used metallodrugs in cancer treatment, presents the important drawback of patient resistance. This resistance is the consequence of different processes including those preventing the formation of DNA adducts and/or their quick removal. Thus, a tool for the accurate detection and quantitation of cisplatin-induced adducts might be valuable for predicting patient resistance. To prove the validity of such an assumption, highly sensitive plasma mass spectrometry (ICP-MS) strategies were applied to determine DNA adduct levels and intracellular Pt concentrations. These two metal-relative parameters were combined with an evaluation of biological responses in terms of genomic stability (with the Comet assay) and cell cycle progression (by flow cytometry) in four human cell lines of different origins and cisplatin sensitivities (A549, GM04312, A2780 and A2780cis), treated with low cisplatin doses (5, 10 and 20 mM for 3 hours). Cell viability and apoptosis were determined as resistance indicators. Univariate linear regression analyses indicated that quantitation of cisplatin-induced G-G intra-strand adducts, measured 1 h after treatment, was the best predictor for viability and apoptosis in all of the cell lines. Multivariate linear regression analyses revealed that the prediction improved when the intracellular Pt content or the Comet data were included in the analysis, for all sensitive cell lines and for the A2780 and A2780cis cell lines, respectively. Thus, a reliable cisplatin resistance predictive model, which combines the quantitation of adducts by HPLC-ICP-MS, and their repair, with the intracellular Pt content and induced genomic instability, might be essential to identify early therapy failure. Significance to metallomicsKnowledge about cisplatin as a chemotherapy drug is extensive, including information about the several processes that contribute to its resistance, the main problem of using this chemical. However, this knowledge and information does not yet allow the early identification of resistant tumors/patients, one of the most desired aims of clinicians. In this work we have developed a model that might allow the early detection of chemical resistance and, more importantly, might do so mostly regardless of the resistance mechanism involved, because it determines the dynamics of adduct induction/repair, considering chemical influx/efflux and induced genomic instability.

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Section: Discussionmentioning

confidence: 99%

Section: Total Intracellular Pt Quantitationmentioning

confidence: 99%

Section: Cisplatin-induced Dna Adduct Quantitationmentioning

confidence: 99%

Section: Cisplatin-induced Dna Adduct Quantitationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Cisplatin resistance in cell models: evaluation of metallomic and biological predictive biomarkers to address early therapy failure

et al. 2017

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Nucleic Acid‐based Electrochemical Sensors Facilitate the Study of DNA Binding by Platinum (II)‐based Antineoplastics

Wu,

Arroyo‐Currás

2024

Angewandte Chemie

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DNA crosslinking agents such as cisplatin and related platinum(II) analogs are effective drugs to treat solid tumors. However, these therapeutics can cause high toxicity in the body, and tumors can develop resistance to them. To develop less toxic and more effective DNA crosslinkers, medicinal chemists have focused on tuning the ligands in square planar platinum(II) complexes to modulate their bioavailability, targeted cell penetration, and DNA binding rates. Unfortunately, linking in‐vitro DNA binding capacity of DNA crosslinkers with their in‐vivo efficacy has proven challenging. Here we report an electrochemical biosensor strategy that allows the study of platinum(II)‐DNA binding in real time. Our biosensors contain a purine‐rich deoxynucleotide sequence, T6(AG)10, modified with a 5’ hexylthiol linker for easy self‐assembly onto gold electrodes. The 3’ terminus is functionalized with the redox reporter methylene blue. Electron transfer from methylene blue to the sensor is a function of platinum(II) compound concentration and reaction time. Using these biosensors, we resolve DNA binding mechanisms including monovalent and bivalent binding, as well as base stacking. Our approach can measure DNA binding kinetics in buffers and in 50% serum, offering a single‐step, real‐time approach to screen therapeutic compounds during drug development.

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Nucleic Acid‐based Electrochemical Sensors Facilitate the Study of DNA Binding by Platinum (II)‐based Antineoplastics

Wu,

Arroyo‐Currás

2024

Angew Chem Int Ed

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Quantitative evaluation of cellular uptake, DNA incorporation and adduct formation in cisplatin sensitive and resistant cell lines: Comparison of different Pt-containing drugs

Cited by 56 publications

References 27 publications

Cisplatin resistance in cell models: evaluation of metallomic and biological predictive biomarkers to address early therapy failure

Cisplatin resistance in cell models: evaluation of metallomic and biological predictive biomarkers to address early therapy failure

Nucleic Acid‐based Electrochemical Sensors Facilitate the Study of DNA Binding by Platinum (II)‐based Antineoplastics

Nucleic Acid‐based Electrochemical Sensors Facilitate the Study of DNA Binding by Platinum (II)‐based Antineoplastics

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