Journal: PLoS ONE

Article Title: Loss of Activating EGFR Mutant Gene Contributes to Acquired Resistance to EGFR Tyrosine Kinase Inhibitors in Lung Cancer Cells

doi: 10.1371/journal.pone.0041017

Figure Lengend Snippet: A, PC9/ER1 cells were treated with or without 1 µM erlotinib, and 5 µM lapatinib for 5 hrs, and followed Western blot analysis. B, PC9/ER1 cells were treated with 10 nM of siRNAs of scrumble and EGFR family genes, and exposed to erlotinib (1 µM) or BIBW2992 (1 µM) for 5 hrs, and followed Western blot analysis.

Article Snippet: Erlotinib was kindly provided by F. Hoffman-La Roche Ltd, gefitinib was by AstraZeneca Inc. BIBW2992 was purchased from Selleck Chemicals, SU11274 and wortmannin were from Calbiochem, LY294002 was from Cell Signaling Technology and Lapatinib was from Toronto Research Chemicals.

Techniques: Western Blot

Journal: Cancers

Article Title: Functional Antagonism of Junctional Adhesion Molecule-A (JAM-A), Overexpressed in Breast Ductal Carcinoma In Situ (DCIS), Reduces HER2-Positive Tumor Progression

doi: 10.3390/cancers14051303

Figure Lengend Snippet: JAM-A antagonism reduces tumor size in a chick embryo xenograft model. ( A ) SUM-225 cells (2 × 10 6 ) were implanted onto the chorioallantoic membrane overlying a developing chick embryo and treated then and 4 days later with 15 µL PBS, 200 µM JBS2, and 0.1 µM lapatinib or JBS + lapatinib. ( A ) Upon xenograft harvesting, the number of grossly visible tumors was observed to be less in treated relative to control conditions according to the following hierarchy: PBS > lapatinib > JBS2 > JBS2 + lapatinib ( n = 5–8 eggs per condition). No embryonic death was observed for JBS2-treated eggs, while there were 1–3 embryonic deaths in the other conditions as follows: PBS < lapatinib < JBS2 + lapatinib (*** p < 0.0001). ( B ) Representative image of a gross xenograft tumor (arrowhead) before harvesting from the CAM. The boundary of the silicon ring is shown as a dashed line. ( C ) Representative images of formalin-fixed paraffin-embedded xenograft tumor sections (4 μm) stained with hematoxylin/eosin.

Article Snippet: These were treated for 28 days as follows: PBS ( n = 8; daily intra-peritoneal/i.p. injection at equivalent volume to JBS2, typically 120–150 μL), JBS2 ( n = 9; 10 mg/kg daily i.p), lapatinib tosylate (Carbosynth Ltd., Berkshire, UK; n = 8; 100 mg/kg oral gavage on 5 consecutive days per week), JBS2 plus lapatinib ( n = 8; 10 mg/kg and 100 mg/kg respectively).

Techniques: Membrane, Control, Formalin-fixed Paraffin-Embedded, Staining

Journal: Cancers

Article Title: Functional Antagonism of Junctional Adhesion Molecule-A (JAM-A), Overexpressed in Breast Ductal Carcinoma In Situ (DCIS), Reduces HER2-Positive Tumor Progression

doi: 10.3390/cancers14051303

Figure Lengend Snippet: JAM-A antagonism reduces tumor progression in an mfp tumor xenograft model. ( A ) SUM-225- luc tumor xenografts grown to a volume of 100 mm 3 in the mfps of NOD/SCID mice were treated for 4 weeks with PBS, JBS2, lapatinib, or JBS2 + lapatinib and volumetrically measured twice/week ( n = 6, 9, 6, 8 mice, respectively). JBS2, lapatinib, and JBS2+lapatinib all significantly reduced tumor growth over time versus PBS-treated tumors (*** p < 0.001). Co-administration of JBS2 with the positive control drug lapatinib significantly shortened the time until lapatinib reached its maximal effect (* p < 0.05, comparing volumes in lapatinib versus JBS2+lapatinib tumors at 1 week post treatment). ( B ) Tumor bioluminescence was imaged once-weekly by in vivo imaging system (IVIS), and mean values over the entire treatment period (weeks 1–4 inclusive) expressed as flux (pixels/second; p/s) were plotted. Graph shows maximum and minimum values with a line at the mean bioluminescence for each treatment group. JBS2 significantly reduced mean tumor bioluminescence (* p < 0.05; PBS versus JBS2 and JBS2 versus JBS2 + Lapatinib). ( C ) FFPE tumor sections were stained with hematoxylin and eosin (top panel) or immunohistochemically assessed for the expression of JAM-A (middle panel) or cytokeratin (CK)-5/6 (lower panel). ( D ) JAM-A expression (measured semi-quantitatively) was reduced in tumor sections from mice treated with JBS2 relative to PBS (* p < 0.05). ( E ) The percentage positivity for cytokeratin 5/6 expression in xenograft tumor cells was not altered between treatment groups ( n = 9–11 mice per group), where ST refers to sub-threshold tumors that never reached 100 mm 3 and were thus not randomized to receive treatment. All treatments reduced the percentage of CK5/6-positive cells in xenograft basement membranes (BM); ( F ) a surrogate for tumor cell invasion.

Article Snippet: These were treated for 28 days as follows: PBS ( n = 8; daily intra-peritoneal/i.p. injection at equivalent volume to JBS2, typically 120–150 μL), JBS2 ( n = 9; 10 mg/kg daily i.p), lapatinib tosylate (Carbosynth Ltd., Berkshire, UK; n = 8; 100 mg/kg oral gavage on 5 consecutive days per week), JBS2 plus lapatinib ( n = 8; 10 mg/kg and 100 mg/kg respectively).

Techniques: Positive Control, In Vivo Imaging, Staining, Expressing

Journal: Cancers

Article Title: Functional Antagonism of Junctional Adhesion Molecule-A (JAM-A), Overexpressed in Breast Ductal Carcinoma In Situ (DCIS), Reduces HER2-Positive Tumor Progression

doi: 10.3390/cancers14051303

Figure Lengend Snippet: JAM-A and HER2 antagonism evoke multiple proteomic changes in mfp tumor xenografts. Fresh-frozen tissues harvested from the SUM-225 mouse mfp model of breast cancer (treatment groups: PBS, JBS2, lapatinib, JBS2+lapatinib) were subjected to proximity ligation assay proteomic analysis using an oncology-centered 92-protein array panel. Seven individual protein targets changed significantly between conditions in the 92-protein array (CAIX, MUC-16, ESM-1, TFPI-2, VEGFA, IL-6, HGF; * p < 0.05).

Article Snippet: These were treated for 28 days as follows: PBS ( n = 8; daily intra-peritoneal/i.p. injection at equivalent volume to JBS2, typically 120–150 μL), JBS2 ( n = 9; 10 mg/kg daily i.p), lapatinib tosylate (Carbosynth Ltd., Berkshire, UK; n = 8; 100 mg/kg oral gavage on 5 consecutive days per week), JBS2 plus lapatinib ( n = 8; 10 mg/kg and 100 mg/kg respectively).

Techniques: Proximity Ligation Assay, Protein Array