Sorafenib

Manufactured by Selleck Chemicals

Sourced in United States, China, Germany, Australia

Sorafenib is a laboratory reagent that functions as a multi-kinase inhibitor. It is commonly used in research settings to study cellular signaling pathways and their modulation.

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Close spelling variants of this product

Sorafenib tosylate (20 citations) Sorafenib (BAY 43-9006) (6 citations) Sorafenib (S7397) (4 citations) Sorafenib (No. S7397), (2 citations)

230 protocols using sorafenib

Sorafenib-resistant Hepatocellular Carcinoma Cells

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The normal human liver cell line LO2 was acquired from the American Type Culture Collection (ATCC). Hepatocellular carcinoma cell lines HepG2 and MMC-7721 were acquired from the China Center for Type Culture Collection. Cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) complemented with 10% fetal bovine serum (FBS) and 100 IU/mL penicillin-streptomycin at 37°C with 5% CO₂. To create the sorafenib-resistant cells (HepG2-R), HepG2 cells were treated with incremental concentrations of sorafenib (Selleck, Houston, TX, USA); starting with 0.5 μM, the concentration was doubled every 2 weeks until it reached 8 μM. Then, the HepG2-R cells were treated with 8 μM of sorafenib biweekly to maintain the resistant ability [14 (link)].
To obtain exosomes, cells were cultured in 15-cm dishes with 30 mL of the whole culture. After reaching 70% confluence, cells were washed with phosphate-buffered saline (PBS) and cultured in DMEM complemented with 10% exosome-depleted FBS for 48 hours. The exosome depleted FBS was obtained by ultracentrifugation at 100 000×g for 12 hours.

Wang G., Zhao W., Wang H., Qiu G., Jiang Z., Wei G, & Li X. (2019). Exosomal MiR-744 Inhibits Proliferation and Sorafenib Chemoresistance in Hepatocellular Carcinoma by Targeting PAX2. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 25, 7209-7217.

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Assessing Sorafenib's Impact on Cell Proliferation

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Sorafenib (Cat No.: S7397) were purchased from Selleck Chemicals and dissolved in DMSO. To assess cell proliferation under the drug treatment, the cells in each condition were seeded at 4 × 10⁴ cells/well in a 12-well plate, 4 × 10⁴ cells/well in a 6-well plate, or 1 × 10⁴ cells/well in a 12-well plate and cultured for 6, 8, or 10 days accordingly in a 37 °C humidified CO₂ incubator with the drug or vehicles containing medium refreshed every other day. The number of cells at day 6, 8, or 10 was determined by crystal violet staining, and their relative absorbance was normalized to the non-targeting control of the same day. To generate Sorafenib-resistant cells, Huh7 cells were treated with either DMSO (vehicle) or 2.5 μM Sorafenib (Selleck Chemicals, S7397) for approximately 4 months. Medium was changed every 3 days.

Zhang Y., Nguyen T.M., Zhang X.O., Wang L., Phan T., Clohessy J.G, & Pandolfi P.P. (2021). Optimized RNA-targeting CRISPR/Cas13d technology outperforms shRNA in identifying functional circRNAs. Genome Biology, 22, 41.

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Sorafenib-Resistant HCC Cell Lines

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Human HCC cell lines (Huh7, SMMC-7721) were obtained from the Cell Bank of the Chinese Academy of Science (Shanghai, China). Huh7 and SMMC7721 cells were cultured in RPMI-1640 containing 10% fetal bovine serum (FBS), 0.1 mg/mL streptomycin and 100 U/mL penicillin and maintained in a 5% CO₂ incubator at 37 °C.
Generation of sorafenib-resistant cells.
Huh7 and SMMC7721 cells were seeded into 25 cm² cell culture flasks. Firstly, when the cell density reached 80%, the cells were cultured in complete medium supplemented with 4.3 μM sorafenib (Selleck, USA) for 48 h. Subsequently, the cells were cultured in sorafenib-free complete medium for a specific duration and then exposed to a medium containing 8 μM sorafenib. After observing approximately 50% cell death, the medium was replaced with sorafenib-free complete medium. Upon steady growth and initiation of proliferation, the cells were cultured in complete medium containing 12 μM sorafenib, and sorafenib was removed after observing approximately 50% cell death. This cyclic culture process was repeated for approximately eight months, following the aforementioned steps. Ultimately, the cells acquired stable resistance to sorafenib and were designated as Huh7-S and SMMC7721-S cells.

Xu K., Wang X., Hu S., Tang J., Liu S., Chen H., Zhang X, & Dai P. (2024). LINC00540 promotes sorafenib resistance and functions as a ceRNA for miR-4677-3p to regulate AKR1C2 in hepatocellular carcinoma. Heliyon, 10(5), e27322.

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Establishment of Sorafenib-Resistant HepG2 Cells

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The HepG2 human hepatoma cell line was purchased from the Shanghai Institute of Biochemistry and Cell Biology (Shanghai, China). All cell lines in the study were maintained in DMEM medium (Gibco, CA, United States) supplemented with 10% fetal bovine serum (Gibco, CA, United States), 50 μg/ml streptomycin, and 100 U/ml sodium penicillin at 37°C in an atmosphere containing 5% CO₂. HepG2 sorafenib-resistant cells (HepG2-SR) were generated by selection starting with sorafenib (Selleckchem, United States) at 2 μm, increasing to 10 μm over the course of 3 months.

He W., Huang X., Berges B.K., Wang Y., An N., Su R, & Lu Y. (2021). Artesunate Regulates Neurite Outgrowth Inhibitor Protein B Receptor to Overcome Resistance to Sorafenib in Hepatocellular Carcinoma Cells. Frontiers in Pharmacology, 12, 615889.

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Sorafenib-resistant Hepatocellular Carcinoma Cell Lines

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HEK-293T, Huh7 and Hep3B cells were obtained from the American Type Culture Collection (ATCC) and cultured at 37°C in an atmosphere containing 5% CO₂ and in DMEM medium (Gibco) supplemented with 10% fetal bovine serum (Gibco). Huh7/sorafenib-resistant (SR) and Hep3B/SR were constructed by long-term exposure to 5 μM sorafenib (Selleck), which was increased to 20 μM over 3 months. 5 μM sorafenib was added to the medium to maintain sorafenib resistance in the Huh7/SR and Hep3B/SR cells.

Liu Y., Chen L., Yuan H., Guo S, & Wu G. (2020). LncRNA DANCR Promotes Sorafenib Resistance via Activation of IL-6/STAT3 Signaling in Hepatocellular Carcinoma Cells. OncoTargets and therapy, 13, 1145-1157.

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Evaluating HBO and Drug Treatments on Hepatoma Cells

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Hepatoma cells were divided into six groups: the blank control group, cells were treated with the vehicle; HBO group, cells were treated with HBO (see HBO treatment); sorafenib group, cells were treated with 1.6 µM sorafenib (Selleckchem, USA); cisplatin group, cells were treated with 1 µM cisplatin (Nanjing Pharmaceuticals, China); HBO + sorafenib group, cells were treated with sorafenib followed by HBO one hour later; HBO + cisplatin group, cell were treated with cisplatin and HBO. Cisplatin, a classical chemotherapy agent that inhibits hepatoma cell growth, was used as a positive control. All treatments were started 24 hrs after cells were seeded in 96-well plates.

Peng H.S., Liao M.B., Zhang M.Y., Xie Y., Xu L., Zhang Y.J., Zheng X.F., Wang H.Y, & Chen Y.F. (2014). Synergistic Inhibitory Effect of Hyperbaric Oxygen Combined with Sorafenib on Hepatoma Cells. PLoS ONE, 9(6), e100814.

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Chemotherapeutic Sensitivity Modulated by miR-93

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We examined the relationship between miR-93 expression and cell sensitivity to two chemotherapeutic drugs for advanced HCC (5-FU and cisplatin) and two tyrosine kinase inhibitors. Three HCC cell lines (HepG2, Hep3B, and PLC/RFP/5) were treated with either anti-miR-93 or control-miR and were divided into 96-well micro plates. To analyze the sensitivity to 5-FU and cisplatin, cells (1×10³) were treated with 0 to 2mM 5-FU (Sigma-Aldrich, St. Louis, MO) or 0 to 1 mM, cisplatin (Santa Cruz Biotechnology, Inc., Dallas, TX). To analyze the sensitivity to sorafenib (Selleck Chemicals, Houston, TX) and tivantinib (Selleck Chemicals), cells (5×10³) were treated with 0 to 50 μM sorafenib or 0 to 25 μM tivantinib. After 24 or 72 hrs, cell viability was determined by the CellTiter-Glo Luminescent Cell Viability Assay Kit (Promega, Fitchburg, WI). All assays were assessed in triplicate.

Ohta K., Hoshino H., Wang J., Ono S., Iida Y., Hata K., Huang S.K., Colquhoun S, & Hoon D.S. (2014). MicroRNA-93 activates c-Met/PI3K/Akt pathway activity in hepatocellular carcinoma by directly inhibiting PTEN and CDKN1A. Oncotarget, 6(5), 3211-3224.

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Establishing Sorafenib-Resistant Hepatocellular Carcinoma Cell Lines

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Human HCC cell lines Huh-7, HepG2, HA59T, HA22T, HCC36, and Mahlavu and sorafenib-resistant cells (Huh-7/SR) were grown in Dulbecco’s modified Eagle medium (Corning, USA) supplemented with 10% fetal bovine serum and 100 U/mL penicillin–streptomycin–amphotericin B (Biological Industries) and were incubated at 37° C in a humidified atmosphere containing 5% CO₂. To induce sorafenib resistance, Huh-7 cells were treated with 0.25 μM sorafenib (BAY 43-9006, Selleck Chemicals, USA), and the dose was gradually increased every week for 6 months. The resistant cells were maintained at a final dose of 10 μM.

Hsu T.W., Su Y.H., Chen H.A., Liao P.H., Shen S.C., Tsai K.Y., Wang T.H., Chen A., Huang C.Y., Shibu M.A., Wang W.Y, & Shen S.C. (2023). Galectin-1-mediated MET/AXL signaling enhances sorafenib resistance in hepatocellular carcinoma by escaping ferroptosis. Aging (Albany NY), 15(13), 6503-6525.

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Investigating miR-518d-5p and Sorafenib Response in Hepatoma Cells

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In vitro experiments were performed using the human hepatoma BCLC3 cell line, obtained and characterized as previously reported^{37 (link)}. The THLE2, Huh7, PLC, and HepG2 cell lines were purchased from ATCC. To carry out in vitro miR-518d-5p inhibition and overexpression, human hepatoma BCLC3 and Huh7 cells were transfected with miRIDIAN microRNA Hairpin Inhibitor/Mimic miR-518d-5p or an unrelated miR-Ctrl using DharmaFECT transfection reagent (Dharmacon, USA) at 25 nM in the culture medium following the manufacturer’s procedure. The study of the response to sorafenib treatment in hepatoma cell lines was carried out using BCLC3 and Huh7 hepatoma cells that were cultured with 10 µM of sorafenib (Selleckchem, USA) for 24 h in a 10% FBS culture medium. Cellular assays were performed at least three times independently. A minimum of three replicates was included per experiment

Fernández-Tussy P., Rodríguez-Agudo R., Fernández-Ramos D., Barbier-Torres L., Zubiete-Franco I., Davalillo S.L., Herraez E., Goikoetxea-Usandizaga N., Lachiondo-Ortega S., Simón J., Lopitz-Otsoa F., Juan V.G., McCain M.V., Perugorria M.J., Mabe J., Navasa N., Rodrigues C.M., Fabregat I., Boix L., Sapena V., Anguita J., Lu S.C., Mato J.M., Banales J.M., Villa E., Reeves H.L., Bruix J., Reig M., Marin J.J., Delgado T.C, & Martínez-Chantar M.L. (2021). Anti-miR-518d-5p overcomes liver tumor cell death resistance through mitochondrial activity. Cell Death & Disease, 12(6), 555.

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Generating Sorafenib-Resistant HCC Cells

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HCC cells with sorafenib resistance were established by long-term exposure to sorafenib (Cat.284461–73-0, Selleckchem, Houston, USA). In detail, HCC cells were treated with low doses of sorafenib (0.625 μM) for 2 weeks, followed by cultivation in a fresh, complete medium for another 2 weeks. Then, the dose of sorafenib was gradually increased and the culture mode was followed successively until the dose of sorafenib was increased to 10 μM, the maximum clinically tolerated dose, and the surviving cells were sorafenib-resistant HCC cells.

Kong H., Sun J., Zhang W., Zhang H, & Li H. (2022). Long intergenic non-protein coding RNA 1273 confers sorafenib resistance in hepatocellular carcinoma via regulation of methyltransferase 3. Bioengineered, 13(2), 3108-3121.

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