TNBC cell lines MDA-MB-231 and SUM159 were purchased from American Type Culture Collection (ATCC), Manassas, VA, USA, and from Asterand, Detroit, MI, USA, These cell lines were chosen based on their high expression of epithelial-mesenchymal transition markers (EMT), metastatic properties, and percentage of CD44+ /CD24- cells. Both cell lines were grown in DMEM (Invitrogen, Carlsbad, CA) with 10 % fetal bovine serum (Cellgro, Manassas, VA) and 1 % penicillin/streptomycin. WST-1 assay was performed using Premixed WST-1 Cell Proliferation Reagent (Clontech, Mountain View, CA) according to the manufacturer’s protocol to access cell viability. Cells were seeded into 96-well tissue culture plates at the concentration of 2,000 cells/well and incubated at different concentrations of Cetuximab (Erbitux, Bristol Myers Squibb, NJ) (0.001–100 μg/ml), Cetuximab (1 nM–100 uM) or in combination with Ixabepilone (Ixampra, Bristol Myers Squibb, NJ) for 72 h. Cell viability was analyzed indirectly by measuring formazan formation by mitochondrial dehydrogenases in viable cells. The absorbance was measured using a multiwall scanning spectrophotometer (Infinite M200 pro, Tecan, Switzerland) at 440 nm (measurement wavelength) and 600 nm (reference wavelength).
Cetuximab
Manufactured by Bristol-Myers Squibb
Sourced in United States
Cetuximab is a monoclonal antibody that targets the epidermal growth factor receptor (EGFR). It is used in the treatment of various types of cancer, including colorectal cancer and head and neck cancer.
Automatically generated - may contain errors
Lab products found in correlation
Trastuzumab, by Genentech (4 mentions)
Everolimus, by Merck Group (2 mentions)
Waters cortecs c18 column, by Waters Corporation (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Prexasertib, by Eli Lilly (2 mentions)
Dmem growth medium, by Merck Group (2 mentions)
Rituximab, by Genentech (2 mentions)
Zeba desalting column, by Thermo Fisher Scientific (2 mentions)
Magnetic polystyrene particles, by Spherotech (2 mentions)
Respective igg isotype control antibodies, by BioLegend (2 mentions)
Cd63 clone h5c6, by BD (2 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Corning (1 mentions)
Sum159, by American Type Culture Collection (1 mentions)
Infinite m200 pro, by Tecan (1 mentions)
Ixabepilone, by Bristol-Myers Squibb (1 mentions)
Premixed wst 1 cell proliferation reagent, by Takara Bio (1 mentions)
Cisplatin, by Bristol-Myers Squibb (1 mentions)
32 protocols using cetuximab
1
TNBC Cell Viability Assays
2
Production and Conjugation of Antibody-Drug Conjugates
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ABT-806 was produced by the transient transfection of HEK-293-6E cells, as described previously [42 (link)]. Maleimidocaproyl (mc) monomethyl-auristatin F (MMAF) and monomethyl-auristatin E (MMAE) to generate ABT-414 and ABBV-221, respectively, were provided by Seattle Genetics. A humanized immunoglobulin-1 (HuIgG1)—conjugated to the cytotoxic payload MMAF or PBD—was used as a negative ADC control. IgG control antibody was produced in-house. Cisplatin and cetuximab (Bristol-Myers Squibb) were purchased. Stock solutions were prepared according to each drug’s specific instructions and stored at −20 °C. Drugs were diluted in fresh media before each experiment.
3
Avelumab and Cetuximab Protocol
4
Cell Line Maintenance and Treatments
5
Characterization of Esophageal and Breast Cancer Cell Lines
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The human esophageal cancer cell lines, KYSE40, KYSE220, KYSE180, and KYSE30 were provided by Dr. Yutaka Shimada34 (link) and have an EGFR expression of 6.0 × 104, 1.3 × 105, 6.0 × 105, and 1.2 × 107 receptors/cell, respectively21 (link). The breast cancer cell lines, MCF-7 and MDA-MD-231 were obtained from ATCC (Manassa, VA, USA). Mouse anti-human EGFR antibodies sc-101 (clone R-1) and sc-120 (clone 528) and mouse anti-human EpCAM antibody sc-59906 (clone HEA125) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Cetuximab (chimeric version of anti-EGFR antibody of clone 225), also known as Erbitux, was obtained from Bristol-Myers Squibb (New York, NY, USA). Goat anti-mouse IgG antibody 1032–01 and rabbit anti-human IgG antibody 6005–1 were purchased from Southern Biotech (Birmingham, AL, USA). Cyanine3 (Cy3) goat anti-mouse IgG antibody CLCC35010 was purchased from Cedarlane (Hornby, ON, Canada). Alexa Fluor 488-labeled donkey anti-mouse IgG antibody A-21202 was purchased from Thermo Fisher Scientific, Inc. (Waltham, MA, USA).
6
Comprehensive Anticancer Agent Evaluation
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Trastuzumab, pertuzumab, and Trastuzumab emtansine were purchased from Roche (Basel, Switzerland). Cetuximab and nivolumab were obtained from Bristol-Myers Squibb (New York, NY, USA). Pembrolizumab was provided by Merck & Co. (Kenilworth, NJ, USA). An anti-Ki-67 antibody (ab16667) was purchased from Abcam (Cambridge, UK).
Seventy-eight anticancer agents were tested in this study (Table S1 ). All compounds were dissolved in dimethyl sulfoxide at a concentration of 20 mM and stored at −80 °C until use. The purity and integrity of the compounds were measured via ultra-performance liquid chromatography-mass spectrometry (Waters Corporation, Milford, MA, USA), using a 1-µL injection volume, as follows. A Waters CORTECS C18 column (particle size: 1.6 µm; column size: 2.1 × 50 mm; Waters Corporation) was developed with a linear aqueous acetonitrile (MeCN) gradient containing a 0.1% formic acid (5–90% MeCN, 1.6 min; flow rate, 1 mL/min), separation was performed at 40 °C, and the components of the major ultraviolet (UV) adsorption peaks were verified by mass spectrometry (Table S1 ).
Epidermal growth factor (EGF) and interferon-γ (IFN-γ) were obtained from Fujifilm Wako Pure Chemical, Ltd. (Osaka, Japan). Staphylococcal enterotoxin B (SEB) was obtained from Sigma-Aldrich (St. Louis, MO, USA).
Seventy-eight anticancer agents were tested in this study (
Epidermal growth factor (EGF) and interferon-γ (IFN-γ) were obtained from Fujifilm Wako Pure Chemical, Ltd. (Osaka, Japan). Staphylococcal enterotoxin B (SEB) was obtained from Sigma-Aldrich (St. Louis, MO, USA).
7
Evaluation of Chk1/2 Inhibitor and Cetuximab in HNSCC
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The HPV-negative UM-SCC1, UM-SCC2, and UM-SCC6 cell lines were obtained courtesy of Dr. Thomas E. Carey (2010; University of Michigan, Ann Arbor, MI). HPV-positive UM-SCC47 and UPCI:SCC090 cells were a gift from Dr. Susan Golin (University of Pittsburgh, Pittsburgh, PA) and Dr. John H. Lee (2011; Sanford Cancer Research Center, Sioux Falls, SD). UM-SCC1-luciferase was obtained from Dr. Eben Rosenthal (2011; Stanford University, Stanford, CA). These cell lines have been previously described (12 (link)–14 (link)). The HPV-negative FaDu (HTB-43) cell line was purchased from the ATCC (2001). UM-SCC1, UM-SCC2, UM-SCC6,UM-SCC47 and UPCI:SCC090 cell lines were maintained in DMEM growth medium (Sigma) supplemented with 10% FBS (SAFC Biosciences) and 1% penicillin/streptomycin (Gibco). The FaDu cell line was maintained in RPMI1640 (Gibco, Invitrogen) supplemented with 10% FBS. The Chk1/2 inhibitor prexasertib (Eli Lilly) was used at 1 nmol/L inUM-SCC1 and UM-SCC47 and at 10 nmol/LinUM-SCC2, UM-SCC6, UPCI:SCC090, and FaDu in vitro, and 4 mg/kg in vivo. Cetuximab (C225, Bristol Myers Squibb) was used at 0.25 μg/mL in vitro and 0.1 mg/injection in vivo.
8
Comprehensive Immunophenotyping of haNK Cells
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The cetuximab (Bristol-Myers Squibb, New York, NY), trastuzumab (Genentech, San Francisco, CA), pertuzumab (Genentech) and isotype (rituximab, Genentech) antibodies were purchased from the National Institutes of Health Pharmacy. Flow cytometry of haNK cells was performed on a BD LSRII flow cytometer (BD Biosciences, San Jose, CA) and analyzed in FlowJo 9.9 (TreeStar Inc., Ashland, OR). Flow cytometry of tumor cells was performed on a BD FACSVerse flow cytometer (BD Biosciences) and analyzed in BD FACSuite (BD Biosciences). Staining of haNK cells was performed with four panels, and the antibodies used were: Anti-CD56-PErCP-Cy5.5, anti-CD16-APC-Cy7, anti-NKG2D-APC, anti-NKG2D-FITC, anti-CD107a-PE-Cy7, anti-GranzymeB-FITC, anti-perforin-APC, anti-NKp46-PE-Cy7, anti-NKp30-APC, anti-NKp44-PE, anti-2B4-FITC, anti-CD158a-PE, anti-CD158d-PE, anti-CD158j-APC, anti-PD-1-PE-Cy7, anti-PD-L1-APC, and anti-MICA-PE; all were obtained from BioLegend (San Diego, CA). Anti-CD56-PE, anti-CD16-PE-Cy7, CD11a-PE, Ki67-FITC, anti-HLA-ABC-FITC and anti-4-1BB-BV421 were obtained from BD Biosciences. In controlled experiments to detect CD16 on haNK cells, the anti-CD16 MAb was CD16-FITC (BD Biosciences) and the isotype control antibody was MIgG1k-FITC (BD Biosciences).
9
Cetuximab Treatment in LGL-KRas Mice
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10
Fluorescent Labeling of Cetuximab
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