Carboplatin resistance was established by deriving Carboplatin-resistant OVCAR3 and HEY-A8 cells from the correspondent parental cells through continuous exposure to Carboplatin (Sigma-Aldrich). OC cells were subjected to initial dose-response studies of Carboplatin (1–100 μM) over 72 h to determine IC50 values. The media was removed and surviving cells were allowed to recover till reached 70–80% confluence. The drug regimen was continued in the same manner for approximately 4–6 cycles. Subsequently, the IC50 concentrations were re-evaluated in the resistant OVCAR3 and HEY-A8 cells. The cells were then continuously maintained in the presence of Carboplatin at these updated IC50 concentrations for an additional 4–6 cycles (23 (link)).
Carboplatin
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Canada, Macao, Australia
Carboplatin is a platinum-based antineoplastic agent used in the treatment of various types of cancer. It functions by inhibiting DNA synthesis and cellular division, thereby disrupting the growth and proliferation of cancer cells.
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Lab products found in correlation
Cisplatin, by Merck Group (49 mentions)
Paclitaxel, by Merck Group (36 mentions)
Doxorubicin, by Merck Group (19 mentions)
Oxaliplatin, by Merck Group (18 mentions)
Etoposide, by Merck Group (14 mentions)
Gemcitabine, by Merck Group (9 mentions)
Docetaxel, by Merck Group (8 mentions)
Olaparib, by Selleck Chemicals (8 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (8 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Pemetrexed, by Merck Group (4 mentions)
Mg132, by Merck Group (4 mentions)
Paclitaxel, by Selleck Chemicals (4 mentions)
Vincristine, by Merck Group (4 mentions)
Camptothecin, by Merck Group (4 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (4 mentions)
Actinomycin d, by Merck Group (3 mentions)
Doxorubicin, by Selleck Chemicals (3 mentions)
Anti β actin antibody, by Santa Cruz Biotechnology (3 mentions)
169 protocols using carboplatin
1
Carboplatin Resistance in Ovarian Cancer
2
Dose-Dependent Impact of Chemotherapy Drugs on Ovarian and Testicular Germ Cells
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Cisplatin and carboplatin (Millipore, UK) were purchased in powder form, made up in distilled water to stock solutions of 1 mg/ml Cisplatin and 10 mg/ml carboplatin. The chemotherapy drugs were added to the culture medium to produce final concentrations that resulted in a dose response for each tissue that ranged from a lowest dose that had no significant effect to a highest dose that affected the health (ovary) or number (testis) of the vast majority of germ cells. The dose response was based upon the health/number of the follicles/germ cells at the end of the culture period. For the ovary, concentrations of 0.5, 1 and 5 μg/ml Cisplatin and 1, 5 and 10 μg/ml carboplatin were added; for the testis, lower concentrations of 0.01, 0.05 and 0.1 μg/ml Cisplatin and 0.1, 0.5 and 1 μg/ml carboplatin were sufficient to produce the required dose response: all doses used were broadly within the range that has been found in patient serum, as detailed below.
3
Paclitaxel and Carboplatin Combination Therapy
4
Preparation and Storage of Chemotherapeutic Agents
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The chemotherapeutic agents, carboplatin (Sigma-Aldrich, St. Louis, MO, United States), doxorubicin (Sigma-Aldrich), vinorelbine (Sigma-Aldrich), toceranib (Sigma-Aldrich), isophosphoramide mustard (MedChemExpress, Monmouth Junction, NJ, United States), and the active metabolite of ifosfamide were used based on previous publications (2–6 ). Zoledronate (Sigma-Aldrich), carboplatin, and vinorelbine were dissolved in sterile distilled H2O at a final concentration of 10 mM. doxorubicin and toceranib were dissolved in DMSO at a final concentration of 10 mM. Stock solutions were aliquoted and kept at −20°C for long-term storage. Isophosphoramide mustard was dissolved in DMSO at a final concentration of 100 mM. The stock solution of isophosphoramide was aliquoted and kept at −80°C. The stock solutions of carboplatin, doxorubicin, vinorelbine, and toceranib were utilized within 1 year after dissolution. Zoledronate and isophosphoramide mustard were used within 6 months following dissolution. Information on the stability of frozen solutions was only accessible for isophosphoramide mustard, and it remains stable within 6 months.
5
Preparation of Carboplatin and Paclitaxel
6
Cisplatin and Carboplatin Cytotoxicity Assay
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For CCK analysis, lentiviral H2030 cells were treated with either 10-70 μM cisplatin (Sigma-Aldrich, USA) or 80-640 μg/ml carboplatin (Sigma-Aldrich, USA) for 24 h. siRNA and miRNA mimics transfected H2030 cells were treated with either 4-32 μM cisplatin or 30-240 μg/ml carboplatin for 48 h. Four thousand cells per well were plated on a 96-well plate in sextuplicate overnight. After drug treatment, cells were then treated with WST-8 Cell Counting reagent (Dojindo, Japan) for 1 h at 37 °C according to the manufacturer’s protocol. Analysis was performed using the Infinite M200 PRO Microplate Reader (Tecan) with 450 nm absorbance and the survival rate was calculated by normalizing untreated cells to 100%.
7
Cell Line Characterization and Drug Evaluation
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Human lung cancer cell lines, A549 and NCI-H358, were purchased from American Type Culture Collection (ATCC, Manassas, VA). A549 and NCI-H358 cells were cultured in RPMI 1640 medium supplemented with 10 % FBS and 1 % antibiotic/antimycotic (v/v) (Life Technologies, Burlington, ON). Cell were maintained in incubators at 37 °C and 5 % carbon dioxide. Cell identity was confirmed through STR profiling (The Centre for Applied Genomics at The Hospital for Sick Children, Toronto, ON).
BMS-754807 was purchased from Sellek Chemicals (Houston, TX). Cisplatin and carboplatin were both purchased from Sigma-Aldrich (Oakville, ON). BMS-754807 was dissolved in DMSO (Sigma-Aldrich, Oakville, ON) and stored at −20 °C while Cisplatin was dissolved in saline and carboplatin was dissolved in water. Cisplatin and carboplatin solutions were made fresh for each experiment, and stored at 4 °C for the duration of the experiment.
BMS-754807 was purchased from Sellek Chemicals (Houston, TX). Cisplatin and carboplatin were both purchased from Sigma-Aldrich (Oakville, ON). BMS-754807 was dissolved in DMSO (Sigma-Aldrich, Oakville, ON) and stored at −20 °C while Cisplatin was dissolved in saline and carboplatin was dissolved in water. Cisplatin and carboplatin solutions were made fresh for each experiment, and stored at 4 °C for the duration of the experiment.
8
Preparation and Dissolution of Nutlin-3a and Carboplatin
9
Genotoxic Drug-Induced Cellular Responses
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Etoposide (Topoisomerase II inhibitor, induces double-strand breaks in genomic DNA) (33419–42-0, Cayman Chemical), Cisplatin (forms intrastrand cross-links with purine bases in genomic DNA) (CAS 15663–27-1, Merck Millipore), Bleomycin (catalyses single-strand as well as double-strand breaks in genomic DNA) (CAS 9041–93-4, Merck Millipore), Doxorubicin (DOXO) (Topoisomerase II inhibitor, induces double-strand breaks in genomic DNA) CAS 25316–40-9, Merck Millipore), Carboplatin (forms intrastrand cross-links with purine bases in genomic DNA) (CAS 41575–94-4, Merck Millipore), Nutlin-3 (MDM2 antagonist, stabilizes p53) (CAS 548472–68-0, Sigma-Aldrich) and Actinomycin D (Intercalates with genomic DNA, inhibits transcription) (CAS 50–76-0, Sigma-Aldrich) were dissolved in DMSO (CAS 67–68-5, AppliChem GmbH) to prepare stock solutions. These were diluted in cell culture media to achieve the indicated final drug concentrations. At the indicated time point post drug treatments, cells were lysed in TRI reagent for RNA extraction or RIPA buffer (50 mM Tris–HCl, pH 8.0, 150 mM NaCl, 1.0% NP-40, 0.5% sodium deoxycholate supplemented with protease and phosphatase inhibitors) for protein extraction.
10
Evaluating Carboplatin Response in HGSOC Organoids
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To test the response of patient‐derived HGSOC organoids to the chemotherapeutic drug carboplatin (Merck), organoids were dissociated into single cells by using enzymatic (TrypLE™, 15 min, 37°C) and mechanical (vortexing and passing through 26G needle) disruption. Subsequently, cells were counted and seeded at 15,000 cells/25 μl Matrigel™ into a 48‐well plate. After maturation of organoids (~ 7–10 days in culture), treatment with carboplatin with a concentration range between 0 and 100 μg/ml was started. At 1 week post‐treatment, cell viability was determined as described above.
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