Journal: Clinical Cancer Research

Article Title: Increased TGF-α as a Mechanism of Acquired Resistance to the Anti-EGFR Inhibitor Cetuximab through EGFR–MET Interaction and Activation of MET Signaling in Colon Cancer Cells

doi: 10.1158/1078-0432.ccr-13-0423

Figure Lengend Snippet: Figure 4. PHA655752 restores sensitivity of GEO-CR and SW48- CR cells to cetuximab. A and C, GEO, GEO-CR (A), SW48, and SW48-CR (C) cells were treated with increased concentrations of PHA665752 (0.01–10 nmol/L) for 96 hours and evaluated for cell proliferation by MTT staining, as described in Materials and Methods. The results are the average SD of three independent experiments, each done in quadruplicate. B and D, GEO-CR (B) and SW48-CR (D) cells were treated with 2 doses of PHA665752 (0.01 or 0.05 nmol/L for GEO-CR and 5 or 10 nmol/L for SW48-CR), with increasing concentrations of cetuximab (0.01–10 mg/mL) or with a combination of both drugs for 96 hours and evaluated for cell proliferation by MTT staining, as described in Materials and Methods. The results are the average SD of three independent experiments, each done in quadruplicate. E and F, GEO-CR (E) and SW48-CR (F) cells were treated with PHA665752, cetuximab, or their combination at the indicated concentrations for 24 hours. The cell lysates were assayed by Western blotting with the indicated antibodies, as described in Materials and Methods. G and H, GEO-CR (G) and SW48-CR (H) cells were treated with PHA665752, cetuximab, or their combination at the indicated concentrations and analysis of in vitro cell migration was performed as described in Materials and Methods.

Article Snippet: PHA665752, a selective MET tyrosine kinase inhibitor (TKI), was purchased from Santa Cruz Biotechnology.

Techniques: Staining, Western Blot, In Vitro, Migration

Journal: Cancer Management and Research

Article Title: Progress toward overcoming hypoxia-induced resistance to solid tumor therapy

doi: 10.2147/CMAR.S58285

Figure Lengend Snippet: Examples of drugs targeting multiple pathways associated with hypoxia and hypoxia inducible factor 1 (HIF-1)

Article Snippet: Signaling , MET kinase , PHA665752 , Funakoshi et al, Puri et al .

Techniques: Activity Assay, Binding Assay

Journal: Frontiers in Molecular Neuroscience

Article Title: Autism Spectrum Disorder Risk Factor Met Regulates the Organization of Inhibitory Synapses

doi: 10.3389/fnmol.2021.659856

Figure Lengend Snippet: Hepatocyte growth factor (HGF) increases gephyrin clustering in vitro . Hippocampal neurons were left untreated or were treated with DMSO, HGF, or PHA-665752. Representative images of gephyrin- or MAP2-stained hippocampal neurons are shown in the absence (A–A”) or presence (B–B”) of HGF. Overview images are shown in ( A,B ; scale bar 20 μm) while insets are enlarged in ( A’,A”,B’,B” ; scale bar 10 μm). Panels (A’,B’) depict confocal images, (A”,B”) processed images for the quantification of gephyrin cluster (red) densities within masks of MAP2-positive dendritic segments (25 μm). ( C) Quantification of gephyrin cluster densities. One-way ANOVA and Tukey’s multiple comparison test F (4,364) = 0.7245. CTR: n = 75, HGF: n = 71, DMSO n = 78, PHA n = 73, PHA/HGF n = 72. *** p < 0.0001, error bars SEM. (D,E) Untreated hippocampal neurons (D) or treated with HGF (E) were stained for phosphorylated p70S6K (pS6K, scale bar 20 μm). Insets are enlarged in ( D’,E’ ; scale bar 10 μm) for determination of mean fluorescence intensities of phosphorylated p70S6K signals within a mask of MAP2-positive cell bodies. Dashed outlines mark the area of the cell bodies. (F) Quantification of phosphorylated p70S6K intensities. CTR n = 57, DMSO n = 83, HGF n = 83, PHA n = 68, PHA/HGF n = 77. One-way ANOVA and Tukey’s multiple comparison test F (4,363) = 3.369. *** p < 0.0001, error bars: SEM.

Article Snippet: Inhibitors were dissolved in dimethylsulfoxide (DMSO), HGF (Merck KGaA, Darmstadt, Germany) in PBS and added to hippocampal neurons (DIV 9) to final concentrations of 50 ng/ml for HGF, 200 nm for mTOR inhibitor rapamycin (Merck), and 1 μM PHA-665752 (Merck KGaA, Darmstadt, Germany).

Techniques: In Vitro, Staining, Fluorescence