The cisplatin and cisplatin + B 1-6-12 groups received cisplatin (Pfizer, USA) diluted in normal saline to the final concentration of 0.5 mg/mL for intraperitoneal injection. The dose of cisplatin was 2 mg/kg twice a week for 5 continuous weeks (20 mg/kg cumulative dose). This dose regimen has been shown to induce peripheral neuropathy in rats [2, (link)28] . The control group received normal saline injection with the volume and schedule equivalent to the cisplatin groups. B1, B6 and B12 (all from Sigma) (100:100:1 by weight) were dissolved in normal saline and given by gavage during the cisplatin treatment once daily for 5 weeks. This ratio of B 1-6-12 was selected based on the previous studies [3, (link)13] (link). Low-dose, medium-dose and high-dose B 1-6-12 groups received 100, 300 and 600 mg/kg/day, respectively.
Cisplatin
Manufactured by Pfizer
Sourced in United States, United Kingdom, Australia, Sweden, Pakistan
Cisplatin is a platinum-based chemotherapy medication used in the treatment of various types of cancer. It is a laboratory-produced compound that functions as an anti-cancer agent.
Automatically generated - may contain errors
Lab products found in correlation
Microplate reader, by Thermo Fisher Scientific (6 mentions)
Paclitaxel, by Pfizer (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (5 mentions)
Epirubicin, by Pfizer (3 mentions)
Gemcitabine, by Pfizer (3 mentions)
Cisplatin, by MedChemExpress (2 mentions)
Irinotecan, by Pfizer (2 mentions)
Glutathione (gsh), by Merck Group (2 mentions)
5 fluorouracil, by Pfizer (2 mentions)
Gsh mee, by Cayman Chemical (2 mentions)
Erastin, by Selleck Chemicals (2 mentions)
N acetyl cysteine (nac), by Merck Group (2 mentions)
Trolox, by Merck Group (2 mentions)
Ferrostatin 1, by Merck Group (2 mentions)
Mannitol, by Merck Group (2 mentions)
Pcna ab29, by Abcam (2 mentions)
P21 antibody 2947, by Cell Signaling Technology (2 mentions)
Alexa 647 conjugated phalloidin antibody, by Thermo Fisher Scientific (2 mentions)
Ri 1 s8077, by Selleck Chemicals (2 mentions)
71 protocols using cisplatin
1
Cisplatin-induced Neuropathy and B1-6-12 Therapy
2
Protective Effects of APS on Cisplatin-Induced Nephrotoxicity
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All animal experiments are in accordance with the appropriate institutional guidelines for animal research, and animal experiments are approved by Shanghai Jiao Tong University School of Medicine (approval no. SYXK2016-0009). 50 mg APS powder was added to 100 ml normal saline to generate APS solution. 8- to 10-week-old male C57BL/6 mice were randomly assigned to 3 groups: control group, n = 5: mice were intraperitoneally injected with normal saline; cisplatin-treated group, n = 5: mice were intraperitoneally injected with 30 mg/kg cisplatin (Hospira, Australia, Pty Ltd); and cisplatin- and APS-treated group, n = 5: mice were intraperitoneally injected with 30 mg/kg APS (MedChem Express) once a day for 10 consecutive days and injected with cisplatin at day 7. Mice were sacrificed after drug injection 72 h via pentobarbital sodium, and their blood and kidney samples were collected for further experiments. The serum creatinine level was determined using an automatic analyzer.
3
Chemotherapy Effects on Tumor Growth
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Cohorts of 10 implanted mice per treatment arm were monitored for tumor growth. Once visible, tumors were measured using metric calipers (Scienceware, cat: 134160001) twice weekly. Mice were sacrificed when tumors reached 15–20 mm on the longest dimension, considered a humane endpoint by the Animal Care Committee. Once tumors reached approximately 300–750 mm3, mice were randomized into control and chemotherapy arms. For single treatment experiments, mouse cohorts were injected once intraperitoneally with 6 mg/kg of cisplatin (1mg/mL, Hospira, DIN:02126613), 9 mg/kg of paclitaxel (2mg/mL, Hospira, DIN:02296624) or 100 mg/kg of 5-fluorouracil (15mg/mL, Hospira, DIN:021827420) while the control group was injected with saline. For combined treatment experiments, the chemotherapy group was treated once with 5.4mg/kg of cisplatin (1mg/mL, Hospira, DIN:02126613) and 9 mg/kg of paclitaxel (2mg/mL, Hospira, DIN:02296624), based on mouse toxicity experiments. Mice were sacrificed 24 hours after the initial treatment (labeled as small tumors) and at the end of the experiment (large tumors). A median of two mice (1–3) per arm were sacrificed for comparisons. Tumors were harvested and saved in OCT and FFPE blocks for molecular/pathological characterization.
4
Cisplatin-Resistant Bladder Cancer Cell Lines
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Three experimental paired sets of two BC cell lines used: a cisplatin-sensitive control cell line T24 (ATCC, Manassas, VA, USA) and a CR cell line T24R2 generated by the treatment of T24 cells with serial dilutions of cisplatin from 0.039 to 40.0 µg/mL [6 (link)10 (link)]. T24R2 cells were resistant to treatment with cisplatin (Pfizer, Seoul, Korea) at concentration up to 2 µg/mL [11 ] and proven significantly more resistant to cisplatin treatment than other cell lines in our previous studies [6 (link)9 (link)10 (link)11 12 (link)]. To measure cell chemosensitivity, exponentially growing cells were incubated with different concentrations of cisplatin (0.1-100 µg/mL) for four days, as previously described [6 (link)9 (link)10 (link)11 ]. All assays were performed in triplicate. Preliminary experiments confirmed that cisplatin at these concentrations had no direct cytotoxic effect during first 24 hours of treatment. The half maximal inhibitory concentrations (IC50) for T24R2 and T24 cell lines were determined treated with 20 µg/mL and 1.25 µg/mL of cisplatin, respectively (not shown in data) [6 (link)10 (link)].
5
Cisplatin-Induced Kidney Injury Model
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The animals were randomly divided into four groups, each containing 10 animals. The control group of rats (C group) received saline 1 ml/day intraperitoneally for 9 days. The cisplatin group (CIS group) received cisplatin (Pfizer PTY. Limited, Bentley, Australia) intraperitoneally in a single dose of 8 mg/kg on the fifth day of experiment. The aminoguanidine group (AG group) was used as a positive control group and received aminoguanidine (Sigma-Aldrich, St. Louis, Missouri, USA) dissolved in physiological saline solution, intraperitoneally, in a dose of 100 mg/kg/24h for 9 days. The cisplatin-aminoguanidine group (CISAG group) of rats received aminoguanidine intraperitoneally in a dose of 100 mg/kg/24h for 9 days and cisplatin intraperitoneally on the fifth day of treatment in a dose of 8 mg/kg.
Ten days after the beginning of the experiment all animals were anaesthetized using 80 mg/kg ketamine (Ketamidor 10%, Richter Pharma AG, Wels, Austria) and sacrificed. Immediately, blood samples were taken from the aorta and the kidneys were subsequently removed.
Ten days after the beginning of the experiment all animals were anaesthetized using 80 mg/kg ketamine (Ketamidor 10%, Richter Pharma AG, Wels, Austria) and sacrificed. Immediately, blood samples were taken from the aorta and the kidneys were subsequently removed.
6
Cytotoxicity Assay of Cisplatin and Paclitaxel
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Cell lines (A2780/A2780cis) and culturing are as previously described.13 (link) Where stated, A2780cis cells were cultured in the absence of Cisplatin. Cisplatin (Hospira, Maidenhead, UK) was donated by the Aseptic Compounding Unit, St. James’s Hospital, Dublin, Ireland. Paclitaxel powder (Sigma, Wicklow, Ireland) was prepared in dimethyl sulphoxide (DMSO). For IC50 analysis, 5000 cells/well in a 96-well plate were treated with concentration ranges of 0.01–800 μM and 9 mg/ml sodium chloride (Cisplatin) and 0.0375–625 nM (Paclitaxel), using vehicle controls of 1 mg/ml mannitol (Cisplatin) and DMSO (Paclitaxel). After 48 h, cell viability was assessed using the Cell Counting Kit-8 (CCK-8, Sigma). IC50 values were calculated using GraphPad Prism (La Jolla, CA, USA).
7
Culturing Cisplatin-Sensitive and Resistant Ovarian Cancer Cell Lines
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The human epithelial serous ovarian cancer cell lines A2780 (cisplatin-sensitive) and A2780cis (cisplatin-resistant) were purchased from the European Collection of Cell Cultures (ECACC, UK) and cultured in a humidified atmosphere at 37 °C, 5 % CO2. They were maintained in RPMI 1640 medium (Sigma, UK) supplemented with 10 % foetal bovine serum (FBS, Lonza, UK), 1 % penicillin/streptomycin mixture (Lonza, UK) and 2 mM Glutamax (Gibco, Biosciences, Ireland). In addition, A2780cis were cultured in 1 μM cisplatin (Hospira, UK) every second passage in accordance with the ECACC guidelines. Cells were regularly checked for signs of bacterial, fungal or mycoplasmal contamination.
8
Cell Viability Assay for Genotoxic Agents
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Cells were seeded into 96 well plates (4000 cells/well) and incubated for 4 hours before treatment with methyl methanesulfonate (MMS) (Sigma-Aldrich, Saint Louis, MO, USA), mitomycin-C (MMC) (Sigma-Aldrich), cisplatin (Hospira) or UVB (Vilber Lourmat, Bio Spectra V5, 312 nm, cells exposed in 100 μL medium, additional 100 μL added after UVB exposure). The drugs were added on day 0, and cells were analyzed 24, 48 and 72 h after UVB treatment and 24, 72 and 96 h after MMC, MMS and cisplatin treatment. MTT (3-(4.5-Dimethylthiazol-2-yl)-2.5 diphenyl-tetrazolium bromide) was added to the cells and OD was measured at 565 nm, and the average from at least 6 wells was used to calculate cell survival.
9
Canine Cisplatin Dosing Protocol
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The time of initiation of cisplatin infusion was measured in hours and defined as t0h (Fig. 1 ). At t0h, the dogs received 18 mg/m2 of cisplatin (Hospira, Leamington Spa, UK). Dogs were weighed on the day preceding the injection and body surface area (m2) calculated using the formula: where constant k = 10.1 and W is the weight of the dog in kilograms [19 ].
The appropriate volume of stock cisplatin solution (1 mg/mL) for each dog was diluted in 0.9% saline to produce a standard volume of 40 mL. The diluted cisplatin solution was administered through the second lumen of the jugular catheter using automatic dispensing syringe at a flow rate of 2.0 mL/min.
The appropriate volume of stock cisplatin solution (1 mg/mL) for each dog was diluted in 0.9% saline to produce a standard volume of 40 mL. The diluted cisplatin solution was administered through the second lumen of the jugular catheter using automatic dispensing syringe at a flow rate of 2.0 mL/min.
10
Cisplatin-Induced Nephrotoxicity: Molecular Mechanisms
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Cisplatin was obtained as commercially used vials from Hospira (Lake Forest, Illinois). Methanol, potassium dihydrogen phosphate, n-butanol, 1,1′,3,3′-tetramethoxypropane, reduced glutathione (GSH), Ellman’s reagent (5,5′-dithio-bis-2-nitrobenzoic acid), thiobarbituric acid (TBA), and trichloroacetic acid (TCA) were purchased from Sigma-Aldrich (St. Louis, Missourie). Kits for the determination of urea, creatinine, and albumin were obtained from Sigma-Aldrich. Superoxide dismutase (SOD) kit was obtained from assay kits (Biodiagnostic, Cairo, Egypt). Rat TNF-α, caspase-9, and caspase-3 enzyme-linked immunosorbent assay (ELISA) kits were purchased from Raybioech (Norcross, Georgia) and Cloud-Clone Corp (Houston, TX), respectively. Immunohistochemistry (IHC) antibodies for NF-κB and cytochrome c were purchased from Thermo Scientific (Waltham, Massachusetts). DNA extraction kit and RNAlater solution were obtained from Qiagen (Hilden, Germany). Other chemicals were of high analytical grade.
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