Azd5363

Manufactured by Apexbio

Sourced in United States

AZD5363 is a selective and potent inhibitor of the serine/threonine protein kinase Akt. It has a molecular formula of C₂₂H₂₃FN₄O₂ and a molecular weight of 394.44 g/mol.

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Lab products found in correlation

3 protocols using azd5363

Comprehensive Breast Cancer Cell Culture Protocol

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MCF7 and SUM149 cells were cultured per American Type Culture Collection (ATCC) recommendations. Primary murine mammary tumor cells were cultured in phenol-free DMEM/F12 (Gibco), with 10% charcoal-stripped FBS (Gibco), 10 μg/ml insulin, and 10 ng/ml EGF. For treatment of estrogen, AZD5363, and 4OHT, tumor cells were cultured in 10% charcoal-stripped FBS in the presence of E2, AZD5363 (ApexBio Technology), 4OHT (Sigma), or dimethyl sulfoxide (DMSO) for the indicated times and then collected for further analysis. To determine cell viability, 50,000 cells were plated in 24-well plates and treated with DMSO or drugs at the indicated concentrations for 5 days. Viable cell numbers were determined on day 1, day 3, and day 5 by an automatic cell counter (Bio-rad) with trypan blue exclusion. For western blot, tissue and cell lysates were prepared as previously reported [19 (link), 23 (link), 37 (link)]. Primary antibodies used are as follows: HSP90 (Santa Cruz), Gapdh (Ambion), ERα (Santa cruz), Brca1, p-Akt (Ser473), p-4E-bp1 (Thr37/46), p-mTor (Ser2248), p-Gsk3β (Ser9), E-cad, Vim, Snail, Slug, p-Fra1 (Ser 265), p-Rb (ser780) (Cell Signaling) and Fn (Abcam).

Wang C., Bai F., Zhang L.H., Scott A., Li E, & Pei X.H. (2018). Estrogen promotes estrogen receptor negative BRCA1-deficient tumor initiation and progression. Breast Cancer Research : BCR, 20, 74.

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Astragaloside IV Modulation in Metabolic Pathways

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Astragaloside IV (purity ≥ 98%) was obtained from Shanghai Forever Biotech Co., Ltd. (Shanghai, China). Astragaloside IV was dissolved in DMSO to prepare 10 mM stock solution and then was diluted at 1,000-fold in culture medium to generate a working concentration at 10 μM. 0.1% DMSO as a solvent control was run concurrently with the experiments. Palmitate (PA, Sinopharm, Shanghai, China) was dissolved in ethanol to prepare 200 mM stock solution and then further diluted with medium containing 10% FFA-free BSA at the ratio of 1:19 to obtain a concentration of 10 mM before use. Metformin was obtained from Sino-American Shanghai Squibb Pharma (Shanghai, China). Akt inhibitor triciribine, MK2206 and AZD5363 was from Apex Bio (Houston, USA). Isoproterenol was from Shanghai Harvest Pharmaceutical Co., Ltd. (Shanghai, China). TNF-α was from R&D Systems.

Du Q., Zhang S., Li A., Mohammad I.S., Liu B, & Li Y. (2018). Astragaloside IV Inhibits Adipose Lipolysis and Reduces Hepatic Glucose Production via Akt Dependent PDE3B Expression in HFD-Fed Mice. Frontiers in Physiology, 9, 15.

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Characterization of Cell Line Models

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The 293T, MCF7, T47D, ZR-75-30 (ZR), BT549, 231, 468, and derived cell lines were from ATCC. The 2FTGH, U1A, U4A, and γ2A cells were described by Watling et al. (25 (link)). All cells were maintained in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal calf serum (Gibco), 100 U/mL penicillin, and 100 mg/mL streptomycin (Sangon Biotech). Antibodies against phospho-JAK2 (Tyr1007/1008), JAK2, phosphor-STAT3 (Tyr705), STAT3, phospho-STAT1 (Tyr-701), STAT1, phospho-STAT5 (Tyr-694), STAT5, phospho-AKT (Tyr-473), AKT, pmTOR, PTEN, p-HER2, HER2, pFGFR, pSRC, pERK, pPKA, pP65, pIKK, IKK, pIκB, IκB, phospho-ERα (Ser167), and ERα were from Cell Signaling Technology. Hygromycin, puromycin, and anti-FLAG were from Sigma-Aldrich. Anti-ZIP was from Abgent. Antibody against mouse IgG was from Santa Cruz. Antibodies against actin and GAPDH were from Goodhere. AZD1480 and ruxolitinib were from SelleckChem. Tamoxifen citrate was from Merck. Transfection reagents were from Qiagen or Invitrogen. The luciferase assay substrate was from Promega. General biochemical reagents were from Sangon Biotech. Restriction, ligation, and PCR enzymes were from Thermo Fisher. The EGFR inhibitor (C3327), the HER2 inhibitor, AG879, the AKT inhibitor, AZD5363, and the NFκB inhibitor, QNZ, were from APExBio.

Zhu N., Zhang J., Du Y., Qin X., Miao R., Nan J., Chen X., Sun J., Zhao R., Zhang X., Shi L., Li X., Lin Y., Wei W., Mao A., Zhang Z., Stark G.R., Wang Y, & Yang J. (2020). Loss of ZIP facilitates JAK2-STAT3 activation in tamoxifen-resistant breast cancer. Proceedings of the National Academy of Sciences of the United States of America, 117(26), 15047-15054.

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