In order to investigate whether apatinib synergizes the anti-cancer effect of docetaxel on A549/DTX cells via regulating autophagy, 10 µM apatinib (Selleck Chemicals) and 10 µM docetaxel (Abmole Bioscience, Inc.) were used alone or in combination to treat the cells at 37°C. After 48 h of incubation, the protein expression levels LC3A, Beclin-1, PARP and p-AKT were determined via western blot analysis, while the apoptosis rate was determined using a Annexin V/PI double staining kit (Calbiochem; Merck KGaA), according to the manufacturer's protocol, in each group of cells.
Apatinib
Manufactured by Selleck Chemicals
Sourced in United States, China
Apatinib is a laboratory reagent used for cell culture and research applications. It is a small molecule tyrosine kinase inhibitor that targets the vascular endothelial growth factor receptor 2 (VEGFR-2). Apatinib is employed in various in vitro and in vivo studies to investigate angiogenesis, tumor growth, and related cellular processes.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Sorafenib, by Selleck Chemicals (2 mentions)
Anlotinib, by Selleck Chemicals (2 mentions)
Dasatinib, by Selleck Chemicals (2 mentions)
Docetaxel, by Selleck Chemicals (2 mentions)
Docetaxel, by AbMole (2 mentions)
Polybrene, by Santa Cruz Biotechnology (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Cck 8 kit, by Dojindo Laboratories (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
N cadherin, by Santa Cruz Biotechnology (1 mentions)
Antibodies to fibronectin, by Abcam (1 mentions)
Hrp conjugated secondary antibodies, by Vazyme (1 mentions)
Epidermal growth factor (egf), by R&D Systems (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
E cadherin, by Abcam (1 mentions)
Tgf β1, by R&D Systems (1 mentions)
Interleukin 6 (il 6), by R&D Systems (1 mentions)
Osimertinib, by Selleck Chemicals (1 mentions)
Dmem media, by Sartorius (1 mentions)
23 protocols using apatinib
1
Apatinib and Docetaxel Synergy in A549/DTX Cells
2
Synergistic Anti-Cancer Effects of Apatinib and Docetaxel
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In order to investigate whether apatinib synergizes the anti-cancer effect of docetaxel in wild-type A549 cells via regulating autophagy, 10 µM apatinib (Selleck Chemicals) and 10 µM docetaxel (Abmole Bioscience, Inc.) were used to treat wild-type A549 cells alone or in combination at 37°C. After 48 h of treatment, the expression levels of LC3A and Beclin-1 were determined via western blot analysis, and the cell apoptosis rate was determined with an Annexin V/PI double staining kit (Calbiochem; Merck KGaA), according to the manufacturer's protocol, in each group of cells.
3
Gastric Carcinoma Cell Line Responses
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The human gastric carcinoma cell line, HGC-27, was obtained from the American Type Culture Collection. Cells were cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (HyClone; Cytiva), 100 U/ml penicillin and 100 U/ml streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.) in an incubator with 5% CO2 at 37˚C. Cell propagation was performed every 24 h. The cells in the logarithmic phase were used for further analysis.
After cells were fully adhered to the well, they were cultured at 37˚C at a density of 2x103 cells/cm2 and divided into the following groups: i) Control group; ii) IL-17 group; iii) IL-17-apatinib group; and iv) apatinib group. Cells in the IL-17 group were treated with 10 ng/ml IL-17 (Invitrogen; Thermo Fisher Scientific, Inc.) for 48 h. The apatinib group was treated with 50 ng/ml apatinib (cat. no. S7297, Selleck Chemicals) for 48 h. For the IL-17-apatinib group, cells were cultured with 10 ng/ml IL-17 and 50 ng/ml apatinib for 48 h. Cells in the Control group were treated with same volume of DMEM. All treatments were performed at 37˚C.
After cells were fully adhered to the well, they were cultured at 37˚C at a density of 2x103 cells/cm2 and divided into the following groups: i) Control group; ii) IL-17 group; iii) IL-17-apatinib group; and iv) apatinib group. Cells in the IL-17 group were treated with 10 ng/ml IL-17 (Invitrogen; Thermo Fisher Scientific, Inc.) for 48 h. The apatinib group was treated with 50 ng/ml apatinib (cat. no. S7297, Selleck Chemicals) for 48 h. For the IL-17-apatinib group, cells were cultured with 10 ng/ml IL-17 and 50 ng/ml apatinib for 48 h. Cells in the Control group were treated with same volume of DMEM. All treatments were performed at 37˚C.
4
Apatinib Modulates PKM2 Expression in Thyroid Cancer Cells
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SW579 and TPC-1 cells were first disposed of 20 ng/ml, 50 ng/ml, 100 ng/ml Apatinib (Selleck Chemicals; cat. no. S5248) for 48 h51 (link). PKM2 overexpression plasmid and empty vector, PKM2 shRNAs and control were provided by Integrated Biotech Solutions (Shanghai, China). SW579 and TPC-1 cells were (density about 60%) in 6-well plate were transfected with PKM2 overexpression plasmid and vector by applying Lipofectamine 3000 (Invitrogen) in accordance with the reagent instructions. PKM2 shRNAs lentivirus was packaged by WZ Bioscience (Shandong, China). And lentivirus was applied to infect SW579 cells at MOI = 50, and then added with 5 mg/ml Polybrene (Santa Cruz). After 96 h of infection, cells were added with 4 μg/ml Puromycin to screen stable expression cells.
5
Evaluating Anti-Osteosarcoma Compounds
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Human Saos-2 and 143b OS cells were obtained from the Chinese Academy of Sciences. Cells were cultured in DMEM containing 10% FBS (Gibco, CA, USA) in a humidified atmosphere containing 5% CO2 at 37℃. DHA, apatinib, anlotinib, and sorafenib were purchased from Selleck Chemicals (Houston, TX, USA). These agents were dissolved in DMSO (Sigma-Aldrich, St. Louis, MO, USA). OS cells were seeded into 96-well plates. After treatment, CCK-8 was added and incubated with cells according to the instructions of the CCK-8 kit (CCK8; Dojindo, Kumamoto, Japan). The absorbance was measured at 450 nm.
6
Characterization of Epithelial-Mesenchymal Transition in Liver Cells
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Following reagents (suppliers) were used in this study: apatinib (Selleck, Texas, USA); hepatocyte growth factor (HGF) (PeproTech, Suzhou, China); antibodies to CD68, ZO-1, N-cadherin, vimentin, Slug and β-actin (Santa Cruz Biotechnology, USA); antibodies to fibronectin, E-cadherin, Snail, p-Met, Met (Abcam, Cambridge, MA). HRP-conjugated secondary antibodies (Vazyme, Nanjing, China). ELISA kit for HGF, EGF, TGF-β1 and IL-6 (R&D Systems, Minneapolis, MN).
7
NSCLC Cell Lines and Targeted Therapies
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The human NSCLC cell lines PC9 (exon 19 deletion) and H1975 (exon 21 L858R and exon 20 T790M) were obtained from the American Type Culture Collection. Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) media (Biological Industries, Kibbutz Beit Haemek, Israel) or RPMI 1640 media (Biological Industries), supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin at 37 °C in 5% carbon dioxide. Osimertinib and apatinib were purchased from Selleck Chemicals (Shanghai, China).
8
In vivo VEGFR2 signaling blockade
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To block VEGFR2 signaling in vivo, mice were intraperitoneally injected with small-molecule inhibitors of VEGFR2 tyrosine kinase, a VEGFR2-blocking antibody, or their respective vehicles, 4.5 days following surgery. Blocking agents were (i) apatinib [median inhibitory concentration (IC50) = 1 nM, 60 mg/kg S2221; Selleckchem] in 50% dimethyl sulfoxide (DMSO) in phosphate buffer (140 mM NaCl, 9 mM Na2HPO4, 1.3 mM NaH2PO4, pH 7.4) (63 (link), 64 (link)); (ii) cabozantinib (IC50 = 0.035 nM, 30 mg/kg, S1119, Selleckchem) (29 (link)); (iii) ZM323881 HCl (IC50 < 2 nM, 60 mg/kg, S2896, Selleckchem) in 2% DMSO, 30% PEG-300 (Sigma-Aldrich), 5% Tween 80 (Sigma-Aldrich), and 63% ddH2O (30 (link)); and (iv) DC101 (40 mg/kg, BE0060, rat IgG1, Bio X Cell) and nonspecific rat IgG1 control antibody (40 mg/kg, BE0088, Bio X Cell) in PBS (pH 7.0) (31 (link), 51 (link)). Intravital microscopy of the EDL muscle was performed 16 to 18 hours after reagent injection. Thick-section confocal microscopy for quantifying pillars and sprouts after in vivo blockade of VEGFR2 was performed on tissues fixed immediately thereafter, using 400 × 400 μm tiled fields of view, with 16-μm depth.
9
Culturing Lung and Fibroblast Cells
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Human bronchial epithelial cell BEAS-2B, human lung cancer cells A549, and the mouse fibroblast cell NIH3T3 were obtained from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). Cells were cultured in DMEM or RPMI1640 supplemented with 10% fetal bovine serum (FBS) at 37°C in a humidified atmosphere with 5% CO2. Apatinib, PD98059 and Dasatinib were purchased from Selleck Chemical (Houston, TX, USA) at an initial concentration of 10 mM, 20 mM and 10 mM (in 1 mL DMSO). Apatinib and PD98059 were directly used without dilution. Dasatinib was diluted to a final concentration of 50 μM. The final DMSO concentration in cell treatment was 0.04%–0.16%.
10
Inhibiting PD-L1 and VEGFR2 in Ovarian Cancer
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PD‐L1 inhibitor (durvalumab) and VEGFR2 inhibitor (apatinib) were obtained from Selleck Chemicals (Houston, TX, USA). OVCA433 and HeyA8 cells were treated with 4 μmol/L durvalumab (A2013), 20 μmol/L apatinib (S5248), and the two‐drug combination for 48 h using transwell and wound healing assays.
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