Sorafenib (Selleck Chemicals, Houston, TX, USA) and bufalin (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) were dissolved in dimethyl sulfoxide (Sigma-Aldrich; Merck KGaA) and diluted to their working concentrations (Sorafenib at concentrations of 2.5, 5, 10 µM and bufalin at concentrations of 5, 10 and 20 nM). Antibodies against Bcl-2 (Abcam, Cambridge, UK; cat. no. ab692), Bax (Abcam; cat. no. ab32503), caspase 7 (Bioworld Technology, Inc., St Louis Park, MN, USA; cat. no. BS6544), caspase 8 (Bioworld Technology, Inc.; cat. no. AP0237), PARP (Bioworld Technology, Inc.; cat. no. BS70001) and GAPDH (Bioworld Technology, Inc.; cat. no. MB001) were also used.
Bufalin
Manufactured by Merck Group
Sourced in United States, Germany
Bufalin is a cardiac glycoside compound extracted from the skin and venom of certain toads. It functions as an inhibitor of the sodium-potassium ATPase enzyme, which plays a crucial role in regulating the sodium and potassium balance within cells.
Automatically generated - may contain errors
Lab products found in correlation
Digoxin, by Merck Group (5 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (4 mentions)
Gapdh, by Cell Signaling Technology (4 mentions)
Sorafenib, by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Digitoxin, by Merck Group (2 mentions)
Sorafenib, by Selleck Chemicals (2 mentions)
Caspase 7, by Bioworld Technology (2 mentions)
Caspase 8, by Bioworld Technology (2 mentions)
Bcl 2, by Abcam (2 mentions)
Bcl2 associated x (bax), by Abcam (2 mentions)
Gapdh, by Bioworld Technology (2 mentions)
β actin, by Cell Signaling Technology (2 mentions)
Bcl 2, by Cell Signaling Technology (2 mentions)
Phospho akt, by Cell Signaling Technology (2 mentions)
Diphenyleneiodonium chloride, by Merck Group (2 mentions)
Rottlerin, by Merck Group (2 mentions)
Alisertib, by Selleck Chemicals (2 mentions)
Jnk in 8, by Selleck Chemicals (2 mentions)
Barasertib, by Selleck Chemicals (2 mentions)
37 protocols using bufalin
1
Sorafenib and Bufalin Cytotoxicity Assay
2
Investigating the Effects of UV Irradiation and Bufalin on Cell Survival
3
Preparation of Drug Stock Solutions
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Bufalin and LY294002 were purchased from Sigma (St. Louis, MO, USA). Sorafenib and perifosine were purchased from Jinan Trio Pharmatech Co., Ltd. (Jinan, China). The Bufalin, Sorafenib and LY294002 were dissolved in 100% dimethyl sulfoxide (DMSO; Sigma, St. Louis, MO, USA) to prepare stock solutions of 1 mM, 100 mM, and 100 mM, respectively, which were diluted in DMEM to the desired concentrations for the in vitro assays. All stock solutions contained a final DMSO concentration of less than 0.1%. Perifosine was dissolved in phosphate-buffered saline to prepare a 30 μM stock solution.
4
Preparation of Stock Solutions
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ProA (Sigma-Aldrich, Saint-Quentin Fallavier, France), digoxin (Sigma-Aldrich), bufalin (Merck, Fontenay-Sous-Bois, France) and digitoxin (Sigma-Aldrich) were dissolved at 18.8, 12.8, 10 and 10 mM respectively, in cell culture-grade dimethyl sulfoxide (DMSO) and stored at −20 °C until use. Vinblastine (Sigma-Aldrich) was solubilized in sterile distilled water. SB216763 (Sigma-Aldrich) was solubilized in DMSO. All these solutions were freshly diluted in the culture medium for experiments.
5
Investigating Stem Cell Markers in Cancer
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Bufalin and dimethyl sulfoxide were obtained from Sigma-Aldrich (St. Louis, MO, USA). Rabbit anti-CD44 polyclonal antibody (1:1,000; cat. no. BS6825) and horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (1:10,000; cat. no. BS13287) were purchased from Bioworld Technology (Minneapolis, MN, USA). Rabbit anti-ESA polyclonal antibody (1:1,000; cat. no. 21050-1-AP) was purchased from Proteintech Group, Inc. (Wuhan, China). Phycoerythrin-conjugated mouse anti-CD44 monoclonal antibody (1:100; cat. no. 130-095-180), allophycocyanin-conjugated mouse anti-CD24 monoclonal antibody (1:100; cat. no. 130-095-954), and fluorescein isothiocyanate (FITC)-conjugated mouse anti-ESA monoclonal antibody (1:100; cat. no. 130-080-301) for flow cytometry were purchased from Miltenyi Biotec GmbH (Bergisch Glad-bach, Germany). Rabbit anti-Patched (PTCH)1 monoclonal antibody (1:1,000; cat. no. 2468), PTCH2 polyclonal antibody (1:1,000; cat. no. 2464), glioma-associated oncogene 1 (Gli1) monoclonal antibody (1:1,000; cat. no. 3538) and GAPDH monoclonal antibody (1:1,000; cat. no. 8884) were purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA).
6
Assessing EOC Cell Chemoresistance
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The method to assess EOC cancer cell in vitro chemoresistance was described in our previous study.23 Briefly, OVCAR‐3 and ES‐2 cells were lifted off from 6‐well plates and equally plated in a 96‐well plate at a concentration of 5000 cells/well. Bufalin (Sigma Aldrich, USA) was added into tested wells at various concentrations of 0, 0.25, 1, 5, and 10 ng/mL for 48 hours. After that, EOC wells were treated with 20 µL of CellTiter 96® AQueous One Solution Reagent (Promega, USA) and examined on a spectraMax M3 multi‐mode microplate reader (Molecular Devices, USA). The relative percentages of healthy EOC cells were calculated by normalizing the absorbance recordings at 490 nm of all tested wells to the absorbance recordings of wells incubated with 0 ng/mL Bufalin.
7
Evaluating Bufalin Cytotoxicity in Cancer Cells
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The CAPAN-2 human pancreatic cancer cell line (HTB-80) and the CAL-27 human oral cancer cell line (CRL-2095) were purchased from the American Type Culture Collection (USA). The CAPAN-2 cells were cultured in RPMI-1640 medium supplemented with 10% FBS. The CAL-27 cells were grown in DMEM supplemented with 10% FBS, 100 U/mL penicillin, and 100 μg/mL streptomycin. Both cell lines were maintained at 37°C and 5% CO2 in a humidified incubator. Cells were treated with bufalin (Sigma-Aldrich Corp., USA) at the indicated concentrations as in each experiment. Buffer solution containing 0.1% DMSO was used as vehicle control.
8
Bufalin-Induced Apoptosis in Leukemia Cells
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Cell lines K562 and K562/A02 cells were purchased from the Shanghai Institute of Cell Sciences, Chinese Academy of Sciences (Shanghai, PRC). ADM and bufalin were purchased from Sigma-Aldrich (St. Louis, MO). Nrf2, HO-1, P-gp, Bax, Bcl-2, cyctc, PAPR, IRE1α, TRAF2, JNK, p-JNK, and Caspase-12 antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX). The apoptosis detection kit was purchased from KeyGen Biotech (Nanjing, PR China). RT-PCR and HiPerfect transfection kits were purchased from Qiagen (Hilden, Germany).
9
Characterization of Cancer Stem Cells
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Cisplatin was purchased from Qilu Pharmaceutical (Jinan, China). Bufalin was purchased from Sigma (St. Louis, MO, USA). CD44 (60224-1-IG), CD133 (18470-1-IG), OCT4 (11263-1-AP), SOX2 (11064-1-AP), and NANOG (14295-1-AP) primary antibodies were purchased from Proteintech (Chicago, IL, USA). GAPDH (#2118) and ABCG2 (#42078) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).
10
Bufalin Modulates CCRK Expression
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HCC cell line PLC5 and normal immortalized liver cell line LO2 were maintained in the high-glucose DMEM media (Gibco) supplemented with 10% fetal bovine serum (FBS; Hyclone). The cells were cultured at 37 °C in 5% CO2-containing humidified incubator. Bufalin was purchased from Sigma and prepared for the store solution by dissolved in DMSO. In the experiments, store solution of Bufalin was diluted with culture media into indicated concentration and incubated with cells. CCRK-expressing vector and short hairpin RNA vector targeting CCRK (shCCRK) were kindly provided by Marie Lin in Chinese University of Hong Kong. Vectors were transfected into cells with lipofactamine 2000 (Invitrogen) according to the manufacturer’s instructions. For the construction of stable CCRK-depriving PLC5 and stable CCRK-expressing LO2 cells, transfected cells were transferred into selective antibiotics and cultured until the growth of antibiotics-resistant colonies.
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