kinase inhibitors lapatinib [di-p-toluenesulfonate salt]  (LC Laboratories)

Journal: Cancers

Article Title: Liver X Receptor Ligand GAC0001E5 Downregulates Antioxidant Capacity and ERBB2/HER2 Expression in HER2-Positive Breast Cancer Cells

doi: 10.3390/cancers16091651

Figure Lengend Snippet: Co-treatment with 1E5 and lapatinib revealed additive inhibitory effects. ( A ) Lapatinib IC 50 was measured following treatments with varying concentrations (1 nM to 100,000 nM). ( B ) Combination treatments with 1E5 (5 μM) and lapatinib (50 nM) exhibited additive effects. The data were collected from three biological replicates ( n = 3). Standard error was plotted in the graphs. Student’s t -test was used to calculate statistical significance (two-tailed, two-sample equal variance), where ** p < 0.01, *** p < 0.001, and not significant (ns) p > 0.05.

Article Snippet: Tyrosine kinase inhibitor lapatinib and fatty acid synthase inhibitor C75 were purchased from MedChemExpress [South Brunswick Township, NJ, USA (Lapatinib #HY-50898, C75 #HY-12364)].

Techniques: Two Tailed Test

Journal: Bioscience Reports

Article Title: Prostasin regulates PD-L1 expression in human lung cancer cells

doi: 10.1042/BSR20211370

Figure Lengend Snippet: ( A ) Immunoblotting of PD-L1 induced by EGF (50 ng/ml for 24 h) in Calu-3 sublines. ( B ) RT-qPCR analysis of PD-L1 expression in Calu-3 sublines treated with EGF (50 ng/ml, n =3, ANOVA P <0.05, * denotes P <0.05 as compared with the vector control (V). ( C ) Top panel, EGF-induced PD-L1 expression in Calu-3P can be inhibited by an EGFR kinase inhibitor (Lapatinib, 0.2 µM) and antibodies against EGFR (Erbitux, 40 µg/ml) and Her-2 (Herceptin, 40 µg/ml); bottom panel, immunoblotting of GAPDH as a loading control. ( D ) Bar graph of (C) ( n =4), * denotes P <0.05 as compared with the control. ( E ) Immunoblotting of EGFR, Alix, CMTM6 and GAPDH. ( F ) Bar graph of EGFR in (E) ( n =3), * denotes P <0.05, as compared with the vector control (V). Abbreviation: RT-qPCR, real-time quantitative polymerase chain reaction.

Article Snippet: The tyrosine kinase inhibitor (TKI) lapatinib was purchased from Santa Cruz Biotechnology (Dallas, TX, Cat. No. sc-202205).

Techniques: Western Blot, Quantitative RT-PCR, Expressing, Plasmid Preparation, Control, Real-time Polymerase Chain Reaction

Journal: Bioscience Reports

Article Title: Prostasin regulates PD-L1 expression in human lung cancer cells

doi: 10.1042/BSR20211370

Figure Lengend Snippet: (Green) Node 1: EGFR signaling, with Erbitux and lapatinib showing effects on prostasin regulation of PD-L1 expression. Node 2: IFNγ signaling via PKCα, with Gö 6976 and U0126 showing effects on the prostasin-mediated potentiation of PD-L1 induction by IFNγ. Node 3: Prostasin and PD-L1 co-localization in exosomes.

Article Snippet: The tyrosine kinase inhibitor (TKI) lapatinib was purchased from Santa Cruz Biotechnology (Dallas, TX, Cat. No. sc-202205).

Techniques: Expressing

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Effect of Combining EGFR Tyrosine Kinase Inhibitors and Cytotoxic Agents on Cholangiocarcinoma Cells

doi: 10.4143/crt.2020.585

Figure Lengend Snippet: Expression of ErbB receptors in surgically resected patient cholangiocarcinoma (CCA) tissue. (A) Absolute RNA expression of epidermal growth factor receptor (EGFR), human epidermal growth factor receptor (HER) 2, HER3, and HER4 transcript levels in paired CCA patient resected tissue and adjacent margin, detected by droplet digital polymerase chain reaction (ddPCR). Significant difference was determined by paired t test. (B) Absolute expression of EGFR, HER2, HER3, and HER4 transcript levels in primary CCA cells isolated from the tumor tissue and primary cells from normal tissue margins, detected by ddPCR. Significant difference was determined by Welch’s t test. *p < 0.05, **p < 0.01, ***p < 0.001 compared with margin. (C) Immunohistochemistry for ErbB proteins in CCA tissues. EGFR: placenta (positive control EGFR); breast cancer (positive control HER2 and HER3) and normal liver (positive control HER4). Immunopositive cells stained with 3,3′-diaminobenzidine (brown) and counterstained with hematoxylin (blue). Asterisks indicate tumor margin. Scale bars=50 μm.

Article Snippet: The proliferative effect of specific ErbB tyrosine kinase inhibitors (afatinib, lapatinib, and erlotinib) (Selleckchem) on human CCA cells was determined by MTT assay.

Techniques: Expressing, RNA Expression, Digital PCR, Isolation, Immunohistochemistry, Positive Control, Staining

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Effect of Combining EGFR Tyrosine Kinase Inhibitors and Cytotoxic Agents on Cholangiocarcinoma Cells

doi: 10.4143/crt.2020.585

Figure Lengend Snippet: Percentage of ErbB stained tumor from the UK and Thai patients

Article Snippet: The proliferative effect of specific ErbB tyrosine kinase inhibitors (afatinib, lapatinib, and erlotinib) (Selleckchem) on human CCA cells was determined by MTT assay.

Techniques: Staining, Immunohistochemistry

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Effect of Combining EGFR Tyrosine Kinase Inhibitors and Cytotoxic Agents on Cholangiocarcinoma Cells

doi: 10.4143/crt.2020.585

Figure Lengend Snippet: The percentage of cell survival of different four cholangiocarcinoma cell lines treated with various concentrations of ErbB inhibitors at 48 hours by MTT assay. Afatinib (A), lapatinib (B), and erlotinib (C). The percentage cell survival of the combined treatments of fixed concentration of gemcitabine (0.1 μM) with afatinib, lapatinib, and erlotinib (0.1, 1, and 10 μM) in HuCCA-1 cells (D) and KKU-M213 (E). *p < 0.05 compared with untreated control group, # p < 0.05 compared with the gemcitabine-treated group The combination effect is described as synergism (blue), or antagonism (red) according to the combination index value.

Article Snippet: The proliferative effect of specific ErbB tyrosine kinase inhibitors (afatinib, lapatinib, and erlotinib) (Selleckchem) on human CCA cells was determined by MTT assay.

Techniques: MTT Assay, Concentration Assay, Control

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Effect of Combining EGFR Tyrosine Kinase Inhibitors and Cytotoxic Agents on Cholangiocarcinoma Cells

doi: 10.4143/crt.2020.585

Figure Lengend Snippet: IC 50 value (μM) of the specific ErbB inhibitors at 48 hours

Article Snippet: The proliferative effect of specific ErbB tyrosine kinase inhibitors (afatinib, lapatinib, and erlotinib) (Selleckchem) on human CCA cells was determined by MTT assay.

Techniques:

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Effect of Combining EGFR Tyrosine Kinase Inhibitors and Cytotoxic Agents on Cholangiocarcinoma Cells

doi: 10.4143/crt.2020.585

Figure Lengend Snippet: Effect of ErbB inhibitors on primary cholangiocarcinoma (CCA) lines in 3D-tumor growth assay (TGA). (A) Four primary cells were derived from surgically resected CCA tissue samples; CCA-UK5, CCA-UK6, CCA-UK7, and CCA-UK9. Primary cells (dashed), matched cancer associated fibroblasts (CAFs) also derived from the same tissue samples (solid), a human mesenchymal cell line (mesenchymal stem cell [MSC] not shown) or a mixture (CAF+cancer cells shown as dotted line) were employed for the 3D-TGA. (B) Growth of cells, CAFs, and primary cells and CAF (red line) or MSC (green line) mixed with the primary cells was measured using AlamarBlue. Percent cell survival of CCA1 and CCA2 treated with gemcitabine/cisplatin (Gem/Cis) (C), erlotinib (D), or afatinib (E). The mean±20% peak serum concentration for patients is given as the dotted and shaded lines (grey).

Article Snippet: The proliferative effect of specific ErbB tyrosine kinase inhibitors (afatinib, lapatinib, and erlotinib) (Selleckchem) on human CCA cells was determined by MTT assay.

Techniques: Growth Assay, Derivative Assay, Concentration Assay

Journal: Frontiers in Oncology

Article Title: HER3 Differentiates Basal From Claudin Type Triple Negative Breast Cancer and Contributes to Drug and Microenvironmental Induced Resistance

doi: 10.3389/fonc.2020.554704

Figure Lengend Snippet: Differential HER3 response to AKT and PI3K inhibitors in Basal and Claudin type TNBC. (A) Biochemical assessment of the effect of AKT (GDC0068) and PI3K (GDC0077) inhibition in a panel of Basal and Claudin type Triple Negative Breast Cancer (TNBC) showing expression and upregulation of total and phosphorylated HER3 and downstream signaling in the Basal type TNBC cells (BT-20, HCC70, and MDA-MB-468). Cells were treated for 48h with the respective kinase inhibitors at 1uM (B) HER3 quantification in the TNBC cell lines tested in A and their corresponding mutational status of PTEN, PI3K, KRAS, and BRAF (C) . (D) Biochemical signalling analysis in response to NRG- 1α and -1β in MDA-MB-468 showing increased signalling with NRG- 1β isoform.

Article Snippet: Cells were seeded into 96-well plates followed by treatment with individual kinase inhibitors GDC0068 (Selleckchem), GDC0077, Gefitinib (Selleckchem), Neratinib (Selleckchem), Lapatinib (LC Laboratories), Sapitinib (FisherScientific), and the HER3 Ab (R&D Systems), in the presence or absence of NRG-1α or -1β.

Techniques: Inhibition, Expressing

Journal: Frontiers in Oncology

Article Title: HER3 Differentiates Basal From Claudin Type Triple Negative Breast Cancer and Contributes to Drug and Microenvironmental Induced Resistance

doi: 10.3389/fonc.2020.554704

Figure Lengend Snippet: Neuregulin but not Hepatocyte Growth Factor decreases sensitivity to kinase inhibitors in Basal type Triple Negative Breast Cancer (TNBC). (A) Viability assay assessing the effect of Neuregulin-1β (50 ng/ml) in the presence of the small molecule inhibitors GDC0068 (1uM) and GDC0077 (1uM) after 96h treatment in a panel of TNBC cell lines in BT-20, HCC-70, MDA-MB-468, BT-549, and MDA-MB-231 and (B) Dose dependent effects of NRG- 1α and -1β isoforms [Low Dose (LD) 5 ng/ml, Medium Dose (MD) 50 ng/ml, and high dose (HD) 100 ng/ml] in the presence or absence of GDC0068 (1uM) or GDC0077 (1uM) for 96h in HCC-70, MDA-MB-468, and MDA-MB-231 TNBC. (C) Effect of HGF treatment (50 ng/ml) after 96h in the presence or absence of GDC0068/GDC0077 at 1uM. Drug response graphs and Heat map graphs show CellTiter-Glo luminescence viability measurements at the end of the experiments compared to untreated control. Experiments were performed in triplicate. Data are means ± SD]. Drug response graphs were analyzed using two-way analysis of variance (ANOVA)/Tukey’s multiple comparison test, *<0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001].

Article Snippet: Cells were seeded into 96-well plates followed by treatment with individual kinase inhibitors GDC0068 (Selleckchem), GDC0077, Gefitinib (Selleckchem), Neratinib (Selleckchem), Lapatinib (LC Laboratories), Sapitinib (FisherScientific), and the HER3 Ab (R&D Systems), in the presence or absence of NRG-1α or -1β.

Techniques: Viability Assay, Control, Comparison

Journal: Frontiers in Oncology

Article Title: HER3 Differentiates Basal From Claudin Type Triple Negative Breast Cancer and Contributes to Drug and Microenvironmental Induced Resistance

doi: 10.3389/fonc.2020.554704

Figure Lengend Snippet: Gefitinib in combination with AKT/PI3K inhibitors sensitizes Basal type Triple Negative Breast Cancer (TNBC). (A) HCC-70, MDA-MB-468, and MDA-MB-231 TNBC cells treated in the presence of NRG-1β (50 ng/ml) does not significantly decrease sensitivity to gefitinib (200 nM) after 96 h. (B) Viability assay of TNBC cells as in (A) treated with the small molecule inhibitors GDC0068 (1uM), GDC0077 (1uM), gefitinib (200 nM), or a combination of the drugs +/- NRG1β (50 ng/ml) for 96h. In the HCC-70 and MDA-MB-468 cell lines, combination therapy demonstrated decreased viability compared to the MDA-MB-231 cell line which showed no significant difference in response, regardless of treatment (C) Biochemical analysis of the p/EGFR and p/HER3 showing decreased but not abolished phosphorylated proteins indicating a potent and sustained signaling in presence of NRG1β in the Basal type TNBC cells, HCC-70, and MDA-MB-231; Cells were treated for 48h at the drug concentrations shown in (B) . The Claudin type MDA-MB-231 cell line showed no HER3 phosphorylation. Drug response graphs show CellTiter-Glo luminescence viability measurements at the end of the experiments compared to untreated control and analyzed using two-way analysis of variance (ANOVA)/Tukey’s multiple comparison test, *<0.05, ***P<0.001]. Experiments were performed in triplicate. Data are means ± SD].

Article Snippet: Cells were seeded into 96-well plates followed by treatment with individual kinase inhibitors GDC0068 (Selleckchem), GDC0077, Gefitinib (Selleckchem), Neratinib (Selleckchem), Lapatinib (LC Laboratories), Sapitinib (FisherScientific), and the HER3 Ab (R&D Systems), in the presence or absence of NRG-1α or -1β.

Techniques: Viability Assay, Phospho-proteomics, Control, Comparison

Journal: Frontiers in Oncology

Article Title: HER3 Differentiates Basal From Claudin Type Triple Negative Breast Cancer and Contributes to Drug and Microenvironmental Induced Resistance

doi: 10.3389/fonc.2020.554704

Figure Lengend Snippet: Pan HER targeting sensitizes Basal type but not Claudin type Triple Negative Breast Cancer (TNBC) cells to AKT and PI3K inhibition. (A) Biochemical assessment of downstream signaling in the PI3K/AKT signaling pathway after combination therapy with neratinib (300 nM) and GDC0068 (1 uM) or GDC0077 (1 uM) after 48 h, showing deceased signaling in the Basal TNBC cell lines but not in the Claudin type TNBC. (B) Viability assay of TNBC cells treated with the small molecule inhibitors GDC0068, GDC0077, neratinib, or a combination of the drugs for 96h [drug treatments as in (A) ]. In the BT-20, HCC-70, and MDA-MB-468 cell lines, combination therapy demonstrated decreased viability compared to the MDA-MB-231and BT-549 cell line which showed no added benefit of combination therapy. Drug response graphs show CellTiter-Glo luminescence viability measurements at the end of the experiments compared to untreated control and analyzed using two-way analysis of variance (ANOVA)/Tukey’s multiple comparison test, *<0.05, **P < 0.01, ***P<0.001]. Experiments were performed in triplicate. Data are means ± SD].

Article Snippet: Cells were seeded into 96-well plates followed by treatment with individual kinase inhibitors GDC0068 (Selleckchem), GDC0077, Gefitinib (Selleckchem), Neratinib (Selleckchem), Lapatinib (LC Laboratories), Sapitinib (FisherScientific), and the HER3 Ab (R&D Systems), in the presence or absence of NRG-1α or -1β.

Techniques: Inhibition, Viability Assay, Control, Comparison