Studies were performed according to USDA regulations under VCU IACUC protocol AD20008. Immuno-competent A/J mice (SAL cell line) or Swiss Webster (Sarcoma 180 cell line) (~20 g) were injected with 5 x 105 cells into their rear flank (10 animals per treatment group for A/J and 8 per group Swiss Webster +/- SD). Tumors were permitted to form for 10 days with tumors at that time exhibiting a mean volume of ~50 mm3-~100 mm3, respectively. For studies with entinostat, mice were treated by oral gavage once every day QD for twenty-one days with vehicle (0.5% carboxymethyl cellulose, Sigma- Aldrich, St Louis MO, 63013, USA) or with axitinib (2 mg/kg). Animals received entinostat (1 mg/kg) on days 1, 4, 8, 12, 15, 18. For studies with valproate, mice were treated by oral gavage once every day QD for twenty days with vehicle (0.5% carboxymethyl cellulose, Sigma- Aldrich, St Louis MO, 63013, USA) or with [axitinib (2 mg/kg) plus sodium valproate (50 mg/kg)]. The body mass and volume of each tumor was assessed every 3-4 days using a digital caliper and tumor volume calculated using the equation volume = (L x W2)/2.
Axitinib
Manufactured by Merck Group
Sourced in United States, Germany
Axitinib is a pharmaceutical product manufactured by Merck Group. It is a lab equipment designed for use in research and development activities. The core function of Axitinib is to inhibit the activity of certain enzymes involved in the process of angiogenesis, which is the formation of new blood vessels.
Automatically generated - may contain errors
Lab products found in correlation
Sunitinib, by Merck Group (3 mentions)
Lapatinib, by Selleck Chemicals (2 mentions)
Sorafenib, by Merck Group (2 mentions)
Rapamycin, by Merck Group (1 mentions)
Fli 06, by Merck Group (1 mentions)
Nucleofector kit, by Lonza (1 mentions)
Afatinib, by Selleck Chemicals (1 mentions)
Dasatinib, by Selleck Chemicals (1 mentions)
Foretinib, by Selleck Chemicals (1 mentions)
Gefitinib, by Selleck Chemicals (1 mentions)
Mycosensor qpcr assay kit, by Agilent Technologies (1 mentions)
Pd173074, by Merck Group (1 mentions)
Opti mem, by Thermo Fisher Scientific (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
Paclitaxel, by Merck Group (1 mentions)
Cisplatin, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by PAN Biotech (1 mentions)
Vegf a, by Thermo Fisher Scientific (1 mentions)
Zm323881, by Selleck Chemicals (1 mentions)
Collagenase 2, by Worthington (1 mentions)
23 protocols using axitinib
2
Notch and VEGF Signaling Modulation
3
EGFR-mutant Lung Cancer Cell Culture
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The human lung cancer cell line PC9 harboring del E746-A750 activating mutation in EGFR was maintained in RPMI1640 supplemented with 10% fetal bovine serum (FBS) and incubated in a humidified atmosphere of 5% CO2 at 37°C. The PC9 cells were kindly provided by Dr. Mayumi Ono (Kyushu University, Fukuoka, Japan) (13 (link), 19 (link), 23 (link)). Cells were routinely confirmed to be free of mycoplasma contamination using mycosensor QPCR Assay kits (Agilent Technologies). Afatinib, lapatinib, foretinib, gefitinib, and dasatinib were purchased from Selleck (Houston, USA). PD173074, cisplatin, paclitaxel and axitinib were from Sigma Aldrich (St. Louis, MO). The construction of pcDNA3-Twist has previously been described (24 (link)). The small interfering RNAs (siRNA) corresponding to FGFR1, Twist1, ZEB1, Snail, and Slug, mRNA and a non-specific siRNA (control) were purchased from Nippon Gene (Tokyo, Japan). Cells were transfected with siRNA duplexes using Lipofectamine RNAiMAX and Opti-MEM (Invitrogen, Carlsbad, CA) according to the manufacturer's recommendations.
4
Murine Melanoma Cell Line Treatments
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Murine melanoma cell lines 4C11− (nonmetastatic) and 4C11+ (metastatic) were cultured in RPMI 1640 medium supplemented with 5% FBS and 1% penicillin (100U·mL−1) and streptomycin (100 μg·mL−1) at 37 °C in 5% CO2 humidified atmosphere. Cell culture reagents were purchased from PAN Biotech (Aidenbach, Germany). Axitinib (PZ0193; Sigma‐Aldrich, St. Louis, MO, USA), a selective inhibitor of VEGF receptors, and 5‐Aza‐2′‐deoxycytidine (5‐Aza‐CdR; Calbiochem, Merck, Darmstadt, Germany) were dissolved in DMSO (PAN Biotech) and stored at a final concentration of 10 mm at −20 °C. 4C11+ cells were treated with different concentrations (40 nm ‐10 μm ) of Axitinib for MTT assay and with 1 μm for all other assays. All treatments were performed for 48 h. 4C11− cells were treated with 10 μm of 5‐Aza‐CdR for 72 h. As a control, cells were treated with the respective volume of DMSO. Final DMSO volume in the cell culture was lower than 0.01%.
5
Fasting, CNO, and Leptin Regulation in hM3Dq^MCH Mice
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Male hM3DqMCH mice at the age of 10-14 weeks were used for this experiment. Mice were fasted starting at 6 p.m. Axitinib (PZ0193, Sigma, 25 mg/kgBW in PEG/H2O 3:7 mixed, PH 2.5) or vehicle (PEG/H2O 3:7 mixed, PH 2.5) were injected i.p. at 5 a.m. (after 11 hr of fasting), CNO (3 mg/kgBW) was injected to all mice at 6 a.m. (after 12 hr of fasting). Leptin (6 mg/kgBW) or saline were injected i.p. at 10 a.m. (after 16 hr of fasting). 30 min after leptin/saline injection, mice were refed and food intake was monitored 1 hr and 4 hr after refeeding. In the following three weeks, mice were given cross-over injections with either vehicle/Axitinib or saline/leptin.
6
Rat Collagen-Based Angiogenesis Assay
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Standard chemicals and reagents, if not stated otherwise, were purchased from SigmaAldrich (Munich, Germany). VEGF-A (PN# 450-32) was purchased from Peprotech (Hamburg, Germany). ZM323881 (PN# S2896) was purchased from Selleckchem (Munich, Germany). Axitinib (PZ0193) was purchased from SigmaAldrich (Munich, Germany). Rat collagen type I (PN# 08-115) was from Millipore (Darmstadt, Germany) and Collagenase II (PN# LS004176) was purchased from Worthington (Lakewood, CA, USA). Opti-MEM I (1x)-GlutaMAX™-I (51985-026) was purchased from Thermo Fisher Scientific.
7
Pharmacological Inhibition of Key Signaling Pathways
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Cells were treated with pharmacological inhibitors for ATM (KU55933, Calbiochem, San Diego, CA, USA; 10 µM) Chk1 (Prexasertib, Selleckchem, Houston, TX, USA; 1 and 3 nM), DNA-PK (NU7026, Selleckchem; 10 µM), EGFR (Tarceva®, Roche, Basel, Switzerland; 10 µM), ER-α (Hydroxy-Tamoxifen, Sigma Aldrich; 10 µM), HER2 (Ontruzant®, MSD SHARP & DOHME GMBH, Haar, Germany; 2 µg/mL and Lapatinib, Selleckchem; 1 µM), MAPK (SB203580, Selleckchem; 10 µM), MDM2 (AMG232, Axon Medchem, Groningen, The Netherlands; 10 µM), MEK (PD98059, Selleckchem; 20 µM), MRNcomplex (Mirin, Sigma Aldrich; 10 µM), PI3K (LY294002, Selleckchem; 10 µM), PARP (Olaparib, Cell Signaling, Frankfurt a. M., Germany; 10 µM), Rad51 (B02, Axon Medchem; 10 µM) and VEGFR (Axitinib, Sigma Aldrich; 1 µM) with the indicated concentrations. Ethanol (for Hydroxy-Tamoxifen), IgG (for Ontruzant) and DMSO (for all other inhibitors) were used as controls.
8
Formulation and Characterization of Cyclodextrin-Based Caffeine-Axitinib Delivery System
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(2-hydroxypropyl)-β-cyclodextrin with an average molar substitution of hydroxypropyl group of 0.67 was purchased from Shandong Binzhou Zhiyuan Biotechnology Co., Ltd. (China). Hydroxypropyl methylcellulose (HPMC, Pharmacoat 2910) was supplied by Wei Ming Pharmaceutical Mfg. CO., Ltd. (Shin-Etsu, Japan). Caffeine was purchased from Siegfried PharmaChemikalien Minden Gmbh (Germany). Glacial acetic acid (Pharma grade) and axitinib were purchased from Sigma-Aldrich (Germany) and Shilpa Medicare Limited (India), respectively. Distilledwater (UNISS, Taiwan) was used for the preparation of all solutions.
9
Modulation of Islet Cell Function
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Isolated islets were used 6 h after isolation and incubated with the VEGFR inhibitor (axitinib, Sigma-Aldrich, 0.1 µM) and/or the IGF-1R inhibitor (Picropodophyllin, PPP, Sigma-Aldrich, 0.1 µM) for 30–60 min prior to incubation with VEGF-A (PeproTech #450-32, 100 ng/mL), IGF-1 (Novus Biologicals 791-MG, 20 ng/mL) or insulin (NovoNordisk, 1 µM) for the following 24 h. After incubation, isolated islets were fixed in 4% PFA for 30 min and immunohistochemistry was performed as described above. Measurements of delta cell filopodia for all experimental groups were performed in a blind fashion, where the person analyzing the images did not know the identity of all experimental groups until the analysis was completed.
10
Axitinib Treatment Regulates Chondrocyte Elimination
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Axitinib (Sigma) was dissolved in DMSO and injected intraperitoneally at a dose of 0.25 mg per animal per day for the C57BL/6 mice for 8 (P21 to P27) or 10 days (P18 to P27 or P21 to P30). The control group of the C57BL/6 mice received the same volume of DMSO injection each day. Transgenic Prx-Cre:GnasR201H mice and corresponding GnasR201H controls were injected daily 0.017 mg per gram of body weight between postnatal 21 and 29 days of age to block the elimination of dead hypertrophic chondrocytes by ingrowing blood vessels. Animals were sacrificed one day after the last Axitinib injection.
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