Azd6244

Manufactured by Selleck Chemicals

Sourced in United States, United Kingdom, China, Germany

AZD6244 is a small molecule inhibitor of the mitogen-activated protein kinase (MAPK) signaling pathway. It specifically targets the MEK enzyme, which is a key component of the MAPK cascade. AZD6244 is commonly used in laboratory research settings to study cell signaling and the role of the MAPK pathway in various biological processes.

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Lab products found in correlation

145 protocols using azd6244

Inhibition of MEK Signaling in Developing Mice

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MAZ51 (Cta# M1695, Sigma-Aldrich, St. Louis, USA) or AZD6244 (Selumetinib, Cat# S1008, Selleckchem, TX, USA) was dissolved in 0.5% DMSO at 1 mg/mL. For in vivo experiments, 10 mg/kg MAZ51, AZD6244 or vehicle was intraperitoneally injected into WT or Pdcd10^LECKO mice from P4 to P14 daily (10 μg/g body weight). Subsequently, mice were analyzed at P15 or P30.

Qin L., Zhang H., Li B., Jiang Q., Lopez F., Min W, & Zhou J.H. (2021). CCM3 loss-induced lymphatic defect is mediated by the augmented VEGFR3-ERK1/2 signaling. Arteriosclerosis, thrombosis, and vascular biology, 41(12), 2943-2960.

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Targeting RICTOR and KRAS in Cancer

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Cells were obtained from American Type Tissue Collection (Manassas, VA) or collaborators, authenticated via STR DNA fingerprinting at UT MD Anderson Characterized Cell Line Core. Whole genome SNP array profiling was used to determine RICTOR amplified (copy number variation (CNV ≥ 3.5) and non-amplified cell lines (CNV ~2) [31 (link)]. Immortalized human bronchial epithelial cells expressing wtKRAS (HBEC3-KT) and KRAS mutant with stable p53 knockdown (HBEC3-KT53KC12) lines were provided by Drs. Gazdar and Minna (UT Southwestern Medical Center, Dallas, TX). Stable RICTOR knockdown was developed using pTRIPZ inducible lentiviral shRNA plasmids (Rictor shRNA #RHS4696, Non-silencing shRNAmir Control (NTC) #RHS4743) (GE Dharmacon, Lafayette, CO). Targeted inhibitors AZD2014 (vistusertib), AZD6244 (selumetinib), and GSK1120212 (trametinib) were obtained from Selleck Chemicals (Houston, TX).

Ruder D., Papadimitrakopoulou V., Shien K., Behrens C., Kalhor N., Chen H., Shen L., Lee J.J., Hong W.K., Tang X., Girard L., Minna J.D., Diao L., Wang J., Mino B., Villalobos P., Rodriguez-Canales J., Hanson N.E., Sun J., Miller V., Greenbowe J., Frampton G., Herbst R.S., Baladandayuthapani V., Wistuba I.I, & Izzo J.G. (2018). Concomitant targeting of the mTOR/MAPK pathways: novel therapeutic strategy in subsets of RICTOR/KRAS-altered non-small cell lung cancer. Oncotarget, 9(74), 33995-34008.

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Melanoma Cell Lines and Compounds

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Melanoma cell lines YUSEEP, YUHIMO, YUWERA, and YUCRATE were obtained from Ruth Halaban (Yale University) in 2020. Melanoma cell lines WM4235, WM4324, WM9, 1205Lu, WM989, WM983B, WM4380 and WM4258, as well as the PDX model WM4223 were obtained from Meenhard Herlyn (Wistar Institute) in 2020. All patient samples were collected under institutional review board (IRB) approval [34 (link)]. Cell lines were tested for Mycoplasma biannually and authenticated using short-tandem repeat fingerprinting. All cell lines are cultured in RPMI-1640 (Corning,10-040-CM) supplemented with 5% fetal bovine serum (FBS; Cytiva, SH30109.03) in the presence of 5% CO₂ at 37 °C. Commercially purchased compounds include palbociclib (SelleckChem, S1116), ribociclib (SelleckChem, S7440), abemaciclib (Apex Biotechnology, A1794), trametinib (SelleckChem, S2673), AZD6244 (SelleckChem, S1008) and VX-11e (SelleckChem, S7709).

Jagirdar K., Portuallo M.E., Wei M., Wilhide M., Bravo Narula J.A., Robertson B.M., Alicea G.M., Aguh C., Xiao M., Godok T., Fingerman D., Brown G.S., Herlyn M., Elad V.M., Guo X., Toska E., Zabransky D.J., Wubbenhorst B., Nathanson K.L., Kwatra S., Goyal Y., Ji H., Liu Q, & Rebecca V.W. (2023). ERK hyperactivation serves as a unified mechanism of escape in intrinsic and acquired CDK4/6 inhibitor resistance in acral lentiginous melanoma. Oncogene, 43(6), 395-405.

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Synthesis and Characterization of Ligand-Metal Complexes

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The synthesis of the ligands and complexes was performed according to the protocol described by Sgarbossa and co-workers [18 (link)]. Complexes and ligands were dissolved in DMSO as a 10 mM stock solution and stored at room temperature. SB202190, AZD6244, and SP600125 were purchased from Selleck Chemical (Houston, TX); they were dissolved in DMSO as a 20 mM stock solution and stored at −20 °C. Vanadyl sulfate (VOSO₄), 4’,6-diamidino-2-phenylindole (DAPI), Mowiol^® 4-88, RNase, propidium iodide, EDTA, protease inhibitor cocktails, and monoclonal anti-β-actin antibody were purchased from Sigma-Aldrich (St. Louis, MO). VOSO₄ was freshly dissolved in medium. DMEM, DMEM-F12, RPMI 1640, and fetal bovine serum (FBS) were purchased from Aurogene (Rome, IT). EGF, insulin, hydrocortisone, penicillin/streptomycin antibiotic mixture, amphotericin B, and glutamine were purchased from Sigma-Aldrich (Milan, IT).

Boscaro V., Barge A., Deagostino A., Ghibaudi E., Laurenti E., Marabello D., Diana E, & Gallicchio M. (2021). Effects of Vanadyl Complexes with Acetylacetonate Derivatives on Non-Tumor and Tumor Cell Lines. Molecules, 26(18), 5534.

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Selective Inhibition of Cancer Pathways

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ABT-199 was generously provided by Abbvie (Chicago, IL, USA). AZD6244, MEK162, SCH772984, SAHA, LBH589, MS275 and FK228 were purchased from Selleckchem (Houston, TX, USA). Tubacin was purchased from Sigma-Aldrich (St. Louis, MO, USA). Stock solutions were made in dimethylsulfoxide, aliquoted and stored at −20°C.

Ramakrishnan V.G., Miller K.C., Macon E.P., Kimlinger T.K., Haug J., Kumar S., Gonsalves W.I., Rajkumar S.V, & Kumar S.K. (2019). Histone deacetylase inhibition in combination with MEK or BCL-2 inhibition in multiple myeloma. Haematologica, 104(10), 2061-2074.

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High-Throughput Screening of Anticancer Agents

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Nocodozole, BEZ235, BKM120, AZD6244 and AKTViii were purchased from Selleck Chemicals LCC. Small interfering RNAs (siRNAs) were from Shanghai Gene Pharma. Dulbecco’s Modified Eagle’s Media (DMEM), Click-iT Alexa Fluor 488 EdU (5-ethynyl-2’-deoxyuridine) imaging kit and Lipofectamine RNAiMAX was purchased from Life Technologies. Foetal Bovine Serum (FBS) was purchased from SAFC Biosciences™, Lenexa, USA. CellTiter 96^® AQueous One Solution Cell Proliferation Assay and Dual-Glo® Luciferase Assay were purchased from Promega Corporation.

Sinha D., Nag P., Nanayakkara D., Duijf P.H., Burgess A., Raninga P., Smits V.A., Bain A.L., Subramanian G., Wall M., Finnie J.W., Kalimutho M, & Khanna K.K. (2020). Cep55 overexpression promotes genomic instability and tumorigenesis in mice. Communications Biology, 3, 593.

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Synergistic Combinatorial Drug Screening

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For the synergy assay, cells seeded the day before were treated with different concentrations of AZD6244 (0.1, 1, 10, 30 and 50 μM, Selleck Chemicals) in combination with NVP-AEW541 (0.1, 0.3, 1, 3 and 10 μM, Cayman Chemical) or LY3009120 (0.1, 0.3, 1, 3 and 15 μM, Selleck Chemicals). The synergy scores were determined using the R package synergyfinder [56 (link)] with the relative growth rates thresholded between 0 and 1 as input (0 meaning no growth or cell death and 1 meaning growth as fast as the DMSO control).

Dorel M., Klinger B., Mari T., Toedling J., Blanc E., Messerschmidt C., Nadler-Holly M., Ziehm M., Sieber A., Hertwig F., Beule D., Eggert A., Schulte J.H., Selbach M, & Blüthgen N. (2021). Neuroblastoma signalling models unveil combination therapies targeting feedback-mediated resistance. PLoS Computational Biology, 17(11), e1009515.

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Breast Cancer Cell Line Treatments and Protein Analysis

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The human breast cancer cell lines MDA-MB-231, BT-549, T47D, MCF-7, BT474, MDA-MB-453 and SKBR3 were cultured in RPMI1640 media with 10% fetal bovine serum (FBS) at 37°C in a 5% CO2 saturated incubator. MDA-MB-231 cells were treated with DMSO, 100nM GSK690693 (AKT inhibitor, Selleck Chemicals, Houston, TX, USA), or 100 nM AZD6244 (MEK inhibitor, Selleck Chemicals, Houston, TX, USA) for 24 hours before harvesting for protein expression analysis.

Tan L., Song X., Sun X., Wang N., Qu Y, & Sun Z. (2016). ART3 regulates triple-negative breast cancer cell function via activation of Akt and ERK pathways. Oncotarget, 7(29), 46589-46602.

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Preparation of Inhibitor Stock Solutions

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Vemurafenib (ChemieTek) was dissolved in dimethyl sulphoxide (DMSO; Sigma-Aldrich) and stored at –20° C at stocks of 100 mM. Leflunomide (Sigma-Aldrich) was dissolved in DMSO and stored at 4° C at stocks of 10 mM. AZD6244 (selumetinib; SelleckChem) was dissolved in DMSO and stored at –20° C at stocks of 2 mM. When aliquots of the stock were in use they were stored at 4° C for no longer than two weeks.

Hanson K., Robinson S.R., Al-Yousuf K., Hendry A.E., Sexton D.W., Sherwood V, & Wheeler G.N. (2017). The anti-rheumatic drug, leflunomide, synergizes with MEK inhibition to suppress melanoma growth. Oncotarget, 9(3), 3815-3829.

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Cell Line Proliferation Assay with AZD6244 and NVP-BEZ235

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The human lung cancer cell line A549 and the human glioblastoma cell line SNB19 were obtained from the “Cell Line Services” company (Heidelberg, Germany) and routinely cultured under standard conditions (37 °C, 5 % CO₂) in Dulbecco’s modified Eagle’s medium supplemented with 10 % FBS, 1 % glutamine and 1 % Penicillin-Streptomycin. For the proliferation assays cells were treated for 24 h with the indicated concentrations of AZD6244 (Selleckchem, Houston, TX, USA) and NVP-BEZ235 (Novartis Institutes for Biomedical Research, Basel, Switzerland) before measurement of the ATP content. For the other experiments of this study cells were treated 16 or 1 h prior to IR with 500 nM AZD6244 or 50 nM NVP-BEZ235, respectively. Drugs were freshly diluted from frozen aliquots stored at −20 °C. Cells treated in parallel with dimethylsulfoxide (DMSO) served as controls.

Kuger S., Flentje M, & Djuzenova C.S. (2015). Simultaneous perturbation of the MAPK and the PI3K/mTOR pathways does not lead to increased radiosensitization. Radiation Oncology (London, England), 10, 214.

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