The cells were incubated with diverse concentrations of EGFR inhibitor lapatinib (Santa Cruz Biotechnology, CAS 231277–92‐2), MET inhibitor crizotinib (Santa Cruz Biotechnology, sc‐3,564,471) or foretinib (Santa Cruz Biotechnology, CAS 849217–64‐7) separately or in combinations (crizotinib/lapatinib and foretinib/lapatinib) for 24 or 48 h. The inhibitors' concentrations used in the experiments (2 or 5 μM crizotinib, 2 or 5 μM foretinib and 5 or 7.5 μM lapatinib) were selected based on previous XTT experiments (data not shown). For each assay, cells were stimulated with 5 nM EGF (Corning) and 30 ng/mL HGF (Sigma Aldrich) to imitate conditions present in the melanoma microenvironment. Cells incubated only with growth factors and 0.1% DMSO (inhibitors solvent) were treated as a control.
Lapatinib
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Lapatinib is a tyrosine kinase inhibitor used in laboratory research. It functions by inhibiting the activity of certain enzymes involved in cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Afatinib, by Santa Cruz Biotechnology (4 mentions)
Neratinib, by Santa Cruz Biotechnology (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Foretinib, by Santa Cruz Biotechnology (2 mentions)
Erlotinib, by Santa Cruz Biotechnology (2 mentions)
Dacomitinib, by Cayman Chemical (2 mentions)
Afatinib, by Boehringer Ingelheim (2 mentions)
Penicillin streptomycin, by Corning (2 mentions)
Dulbecco modified eagle medium (dmem), by Corning (2 mentions)
Crizotinib, by Santa Cruz Biotechnology (1 mentions)
Hepatocyte growth factor (hgf), by Merck Group (1 mentions)
Epidermal growth factor (egf), by Corning (1 mentions)
Rhtnfα, by BD (1 mentions)
Rhifnγ, by BD (1 mentions)
Trastuzumab, by Genentech (1 mentions)
Poziotinib, by Selleck Chemicals (1 mentions)
Ibrutinib, by Selleck Chemicals (1 mentions)
Prism 9, by GraphPad (1 mentions)
Oxaliplatin, by Merck Group (1 mentions)
Trastuzumab, by Roche (1 mentions)
18 protocols using lapatinib
1
Melanoma Cell Response to EGFR, MET Inhibitors
2
Modulation of HER2-Expressing Cell Responses
3
Cytotoxicity Assays Investigating ERBB4 Inhibitors
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Ba/F3 cells (20,000 cells/well) expressing ERBB4 variants were plated on 96-well plates in RPMI1640 medium containing 10 ng/mL NRG-1. Vector control cells were plated in RPMI1640 medium containing 5% WEHI conditioned medium. The cells were incubated in the presence or absence of 0.0005–10 µmol/L erlotinib (Santa Cruz Biotechnology), afatinib (Boehringer Ingelheim), neratinib (SantaCruz Biotechnology), dacomitinib (Cayman Chemicals), poziotinib (Selleck Chemicals), ibrutinib (Selleck Chemicals), or lapatinib (Santa Cruz Biotechnology), for 72 hours before measuring the viability of cells with MTT assay (described above). Sigmoidal dose–response curves were fitted using asymmetric five-parameter non-linear regression in GraphPad Prism 9 and are graphically displayed with GraphPad Prism 9. The IC50 values are calculated with the help of the R package “drc” (34 (link)) by using quadruplicate measurements from the MTT assay for the indicated concentrations from 3–5 individual experiments and fitting four-parametric log-logistic models (LL.4, LL2.4; R). The unpaired t test (Welch two-sample t test) was used to compare IC50 values.
4
Therapeutic Agents Combination Protocol
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Trastuzumab, pertuzumab, and T-DM1 (Roche, Penzberg, Germany) were obtained by the local pharmacy of the University Medical Center in Goettingen. Small-molecule inhibitors afatinib and lapatinib were purchased from Santa Cruz (Dallas, TX), and 5-FU and oxaliplatin from Sigma (Munich, Germany).
5
Culturing HER2+ Breast Cancer Cell Lines
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The BT474 cell line was maintained in Dulbecco modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. The SKBR3 cell line was maintained RPMI1640 supplemented with 10% FBS and 1% penicillin/streptomycin. All cell lines were cultured in humidified incubation at 37 °C with 5% CO2. Lapatinib was purchased from Santa Cruz (Dallas, TX, USA), and 17DMAG-HCL was purchased from Selleck Chemicals (Houston, TX, USA). Stock solutions were prepared with dimethyl sulfoxide (DMSO) and stored at −20 °C. Lapatinib and 17DMAG for animal experiments were purchased from LC laboratories. Lapatinib was dissolved in water with 0.5% carboxymethylcellulose, 1.8% sodium chloride, and 0.4% Tween 80. 17 DAMG was dissolved in water with 0.8% NaCl.
6
Maintenance and Hormone Deprivation of Breast Cancer Cell Lines
7
Synthesis and Characterization of IDF-11774
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IDF-11774 were synthesized as described previously.18 (link) Sunitinib and 5-FU were obtained from Sigma-Aldrich (St. Louis, MO, USA). Sorafenib and lapatinib were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Stock solutions of compounds were prepared in DMSO at 10 mM and stored at −20 °C. The following antibodies were used: anti-β-actin (sc-47778), anti-ACCα (sc-30212), and anti-AMPKα (sc-25792) from Santa Cruz Biotechnology; anti-HER2 (2242), anti-mTOR (2983), anti-p-mTOR (2971), anti-pACC (3661), anti-4EBP1 (9644), anti-p-4EBP1 (9459), and anti-p-AMPK (2531) from Cell Signaling Technology (Beverly, MA, USA); anti-HIF-1α (610958) and anti-cyclin D1 (554180) from BD Biosciences (San Diego, CA, USA); anti-β-tubulin (ab15568) from Abcam (Cambridge, MA, USA).
8
Inhibition of ErbB Kinase Activity in Embryos
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To block ErbB kinase activity in the embryos, 10 µM of tyrosine kinase inhibitors AG 1478 (Calbiochem), erlotinib, gefitinib, lapatinib, afatinib, neratinib, canertinib (all from Santa-Cruz Biotechnologies), osimertinib, and dacomitinib (both from Cayman Chemical), diluted in DMSO, or DMSO alone was applied in E3 medium at 8 hpf, except as indicated for experimentation shown in Figure 4 D. The final concentration of DMSO was 0.1% of the culture medium.
9
Apoptosis and Autophagy Assays
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Equal numbers of cells were plated and treated the following day by serum deprivation for 24 h or with 100 nM lapatinib (Santa Cruz) for 24 h prior to analysis by Caspase-3 activity assay (Sigma) or trypan blue exclusion and cell counting, which was performed using the cellometer mini cell counter (Nexcelom). For autophagy, cells were treated with 100 uM chloroquine (Sigma) for the indicated times and analyzed by Western blotting or microscopy (Nikon E800).
10
Culturing Cell Lines for Drug Testing
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All cell lines were kept at 37° C and 5% CO2. 293T cells and JIMT-1 cells were grown in DMEM (Corning) supplemented with 10% fetal bovine serum (FBS, Gibco). SMF cells were grown in DMEM (Corning) supplemented with 10% FBS and 5 mg/ml insulin (Gemini Bio-Products). All media contained 2 mM glutamine (Corning) and penicillin/streptomycin (Corning), unless stated otherwise. Lapatinib was purchased from Santa Cruz Biotech. Axitinib, Dovitinib, KW-2449, staurosporine, UCN-01, midostaurin, lestaurtinib, PHA-665752, SU-14913, and sunitinib were purchased from Selleck Chemicals.
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