Journal: Clinical Cancer Research

Article Title: Crosstalk to Stromal Fibroblasts Induces Resistance of Lung Cancer to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors

doi: 10.1158/1078-0432.ccr-09-1001

Figure Lengend Snippet: Fig. 1. Fibroblast-derived HGF induces gefitinib resistance in lung cancer cells. A, coculture with fibroblasts induces gefitinib resistance in lung cancer cells. The PC-9 and HCC827 lung cancer cell lines were cocultured with MRC-5 cells or HGF (20 ng/mL), with control IgG or anti-HGF neutralizing antibody (1 μg/mL), in the presence or absence of gefitinib (0.3 μmol/L) for 72 h, and lung cancer cell growth was determined with a Cell Counting Kit-8. *, P < 0.01 (one-way ANOVA). B, HGF production by lung cancer (PC-9 and HCC827), fibroblasts (MRC-5 and IMR-90), and endothelial (HMVEC and HUVEC) cell lines and by primary cultured fibroblasts from tumors of 5 lung cancer patients. Cells were incubated in medium for 48 h, culture supernatants were harvested, and HGF concentrations were determined by ELISA. C, colocalization of fibroblasts and HGF in tumor tissue from a lung cancer patient. Frozen tumor specimens were subjected to triple-color immunofluorescence analyses using the combination of anti–type I collagen antibody (green), anti-HGF antibody (red), and 4′,6-diamidino-2-phenylindole (blue) as described in Materials and Methods. The fluorescent images were digitally merged. Representative result from lung cancer patient 2. Original magnification, ×200.

Article Snippet: The EGFR mutant human lung adenocarcinoma cell lines PC-9 (del E746_A750) and HCC827 (del E746_A750) were purchased from Immuno-Biological Laboratories and the American Type Culture Collection, respectively.

Techniques: Derivative Assay, Control, Cell Counting, Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay, Immunofluorescence

Journal: Clinical Cancer Research

Article Title: Crosstalk to Stromal Fibroblasts Induces Resistance of Lung Cancer to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors

doi: 10.1158/1078-0432.ccr-09-1001

Figure Lengend Snippet: Fig. 3. Fibroblast supernatants induce gefitinib resistance in lung cancer cells. A, supernatants of human fibroblast cell lines, MRC-5 and IMR-90, induce gefitinib resistance in PC-9 and HCC827 cells. Lung cancer cells were incubated for 72 h with or without gefitinib (0.3 μmol/L) in the presence of culture supernatants of MRC-5 or IMR-90 cells (ratio of 1:2) with or without 1 h pretreatment with control IgG or anti-HGF neutralizing antibody (1 μg/mL). Lung cancer cell growth was determined by MTT assays. *, P < 0.01 (one-way ANOVA). B, supernatants of primary cultured fibroblasts from lung cancer patients (PF) induce gefitinib resistance in lung cancer cells. Lung cancer cells were incubated for 72 h with or without gefitinib (0.3 μmol/L) in the presence of culture supernatants of PF1 or PF2 (ratio of 1:2) with or without 1 h pretreatment by control IgG or anti-HGF neutralizing antibody (1 μg/mL). Lung cancer cell growth was determined by MTT assays. *, P < 0.01 (one-way ANOVA). C, HGF induces lung cancer cell resistance to erlotinib. PC-9 cells were incubated for 72 h with various concentrations of erlotinib and/or HGF (20 ng/mL), and cell growth was determined by MTT assays.

Article Snippet: The EGFR mutant human lung adenocarcinoma cell lines PC-9 (del E746_A750) and HCC827 (del E746_A750) were purchased from Immuno-Biological Laboratories and the American Type Culture Collection, respectively.

Techniques: Incubation, Control, Cell Culture

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Quercetin Inhibits the Proliferation and Metastasis of Human Non-Small Cell Lung Cancer Cell Line: The Key Role of Src-Mediated Fibroblast Growth Factor-Inducible 14 (Fn14)/Nuclear Factor kappa B (NF-κB) pathway

doi: 10.12659/MSM.920537

Figure Lengend Snippet: Administration of Que suppressed the proliferation potential of HCC827 cells. For MTT assays, cells were incubated with Que of 25 μM (Que L), 50 μM (Que M), and 100 μM (Que H) for 96 hours. Every 24 hours, cells were subjected to the testing. For colony formation assays, cells were cultured in medium containing 100 μM Que for 2 weeks. ( A ) Detection results of MTT assays. ( B ) Detection results of colony formation assays. * P <0.05 versus the Control group.

Article Snippet: Human NSCLC cell lines HCC827 (7-1150) and NCI-H1650 (7-1031) were obtained from Chi Scientific (MA, USA).

Techniques: Incubation, Cell Culture, Control

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Quercetin Inhibits the Proliferation and Metastasis of Human Non-Small Cell Lung Cancer Cell Line: The Key Role of Src-Mediated Fibroblast Growth Factor-Inducible 14 (Fn14)/Nuclear Factor kappa B (NF-κB) pathway

doi: 10.12659/MSM.920537

Figure Lengend Snippet: Administration of Que inhibited the invasion and migration potentials, and the signaling transduction of Src/Fn14/NF-κB pathway in HCC827 cells. For scratch assays, cells were incubated with Que of 100 μM (Que H) for 48 hours. For Transwell assays, cells were incubated with Que of 100 μM (Que H) for 24 hours. For western blotting assays, cells were incubated with Que of 100 μM (Que H) for 24 hours. ( A ) Detection results of scratch assays. ( B ) Detection results of Transwell assays. ( C ) Detection results of western blotting assays. * P <0.05 versus the Control group.

Article Snippet: Human NSCLC cell lines HCC827 (7-1150) and NCI-H1650 (7-1031) were obtained from Chi Scientific (MA, USA).

Techniques: Migration, Transduction, Incubation, Western Blot, Control

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Quercetin Inhibits the Proliferation and Metastasis of Human Non-Small Cell Lung Cancer Cell Line: The Key Role of Src-Mediated Fibroblast Growth Factor-Inducible 14 (Fn14)/Nuclear Factor kappa B (NF-κB) pathway

doi: 10.12659/MSM.920537

Figure Lengend Snippet: Overexpression of Src blocked the anti-NSCLC function of Que in HCC827 cells. Cells transfected with negative control vector (NC) or Src expression vector (Src) were treated with Que of 100 μM (Que H) and subjected to MTT assay for 96 hours (cells were collected every 24 hours), and colony formation was assay at 2 weeks, Transwell assay at 24 hours, and scratch assay at 48 hours. ( A ) Detection results of Src level. ( B ) Detection results of MTT assays. ( C ) Detection results of colony formation assay. ( D ) Detection results of Transwell assay. ( E ) Detection results of scratch assay. # P <0.05 versus Que H group.

Article Snippet: Human NSCLC cell lines HCC827 (7-1150) and NCI-H1650 (7-1031) were obtained from Chi Scientific (MA, USA).

Techniques: Over Expression, Transfection, Negative Control, Plasmid Preparation, Expressing, MTT Assay, Transwell Assay, Wound Healing Assay, Colony Assay

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Quercetin Inhibits the Proliferation and Metastasis of Human Non-Small Cell Lung Cancer Cell Line: The Key Role of Src-Mediated Fibroblast Growth Factor-Inducible 14 (Fn14)/Nuclear Factor kappa B (NF-κB) pathway

doi: 10.12659/MSM.920537

Figure Lengend Snippet: Que inhibited the growth and metastasis potential of NSCLC in vivo in by inhibiting Src signaling. Mice were injected with different HCC827 cells and administrated with Que of 100 mg/kg body weight for 3 weeks. ( A ) Detection results of tumor volume. ( B ) Detection results of hematoxylin and eosin detection of tumor tissue. ( C ) Detection results of western blotting detection of E-cadherin and N-cadherin. * P <0.05 versus Que H+Src group. Scale bar, 100 μm.

Article Snippet: Human NSCLC cell lines HCC827 (7-1150) and NCI-H1650 (7-1031) were obtained from Chi Scientific (MA, USA).

Techniques: In Vivo, Injection, Western Blot

Journal: Molecular Cancer Therapeutics

Article Title: Proanthocyanidins Inhibit In vitro and In vivo Growth of Human Non–Small Cell Lung Cancer Cells by Inhibiting the Prostaglandin E2 and Prostaglandin E2 Receptors

doi: 10.1158/1535-7163.mct-09-0638

Figure Lengend Snippet: Figure 1. In vitro treatment of human NSCLCs with GSPs inhibits the proliferation potential and induces apoptosis in a dose-dependent manner. A, comparative dose-dependent effect of GSPs on the proliferation potential of human NSCLC cells, A549, H1299, H226, H460, and H157, 48 h after treatment (29). B, the concentration- and time-dependent effect of GSPs on the proliferation potential of normal human bronchial epithelial cells (BEAS-2B). Cell viability was determined using the MTT assay as described in Materials and Methods and is expressed in terms of percent of vehicle-treated (0.2% DMSO) control cells; points, mean of eight replicates; bars, SD. C and D, GSPs induce apoptosis in A549 and H1299 cells. Apoptotic cell death was analyzed as detailed in Materials and Methods. C, the lower-right (LR) quadrant of the fluorescence-activated cell sorting histograms indicates the percentage of early apoptotic cells, and the upper-right (UR) quadrant indicates the percentage of late apoptotic cells. D, total percentages of apoptotic cells in A549 and H1299 are summarized; columns, mean from three experiments; bars, SEM. Significant difference versus non–GSP-treated controls: *, P < 0.05; ¶, P < 0.01; †, P < 0.001.

Article Snippet: The human NSCLC lines (A549, H1299, H226, H460, H1975, H1650, HCC827, and H157) were purchased from the American Type Culture Collection.

Techniques: In Vitro, Concentration Assay, MTT Assay, Control, Fluorescence, FACS

Journal: Molecular Cancer Therapeutics

Article Title: Proanthocyanidins Inhibit In vitro and In vivo Growth of Human Non–Small Cell Lung Cancer Cells by Inhibiting the Prostaglandin E2 and Prostaglandin E2 Receptors

doi: 10.1158/1535-7163.mct-09-0638

Figure Lengend Snippet: Figure 6. A, dietary GSPs inhibit the growth of A549 and H1299 non–small cell lung cancer cells (NSCLC) grown as xenografts in athymic nude mice (29). Tumor xenografts tissues were harvested at the termination of the experiment, and the wet weight of the tumor/mouse in each group is reported in grams as mean SD, n ¼ 10. Statistical significance versus non-GSP–treated controls, , P < 0.05; {, P < 0.01; †, P < 0.001. B, tumor xenograft tissues from control and GSP-treated (0.5%, w/w) mice were used for the analysis of the levels of COX-2 and PGE2 receptors using Western blotting. Dietary GSPs inhibit the levels of COX-2 and PGE2 receptors, EP1, EP3 and EP4, in the tumor xenograft tissues grown in athymic nude mice compared with control tumor xenograft tissues. Representative blots from A549 or H1299 xenografts are presented from theindependent analysis of tumors from 6 animals pergroup with identical results. The relative density (arbitrary) of each band after normalization for b-actin is shown under each immunoblot as the fold change compared with non-GSP–treated control, which was assigned an arbitrary unit 1 in each case. PGE2 was determined in tumor xenograft tissue samples using a PGE2 immunoassay kit following the manufacturer's instructions. The concentration of PGE2 is expressed in terms of pg/mg protein as a mean SD, n ¼ 10. Significantly lower versus non-GSP–treated controls. †, P < 0.001.

Article Snippet: The human NSCLC lines (A549, H1299, H226, H460, H1975, H1650, HCC827, and H157) were purchased from the American Type Culture Collection.

Techniques: Control, Western Blot, Concentration Assay

Journal: Scientific Reports

Article Title: Implementation and Quality Control of Lung Cancer EGFR Genetic Testing by MALDI-TOF Mass Spectrometry in Taiwan Clinical Practice

doi: 10.1038/srep30944

Figure Lengend Snippet: Accuracy Test of MALDI-TOF MS in EGFR Mutation Testing by Traceable Biological Materials.

Article Snippet: HCC827 , Human lung adenocarcinoma cell line , ATCC , Del19 , No , 0.0 , No , 0.1 , Yes , 95.5 , ATCC CRL-2868.

Techniques: Mutagenesis, Plasmid Preparation, Clone Assay, Expressing

Journal: International Journal of General Medicine

Article Title: Identification of Prognostic Factors Related to Super Enhancer-Regulated ceRNA Network in Metastatic Lung Adenocarcinoma

doi: 10.2147/IJGM.S332317

Figure Lengend Snippet: Identification of super enhancers in LUAD cells. ( A ) Enhancers were ranked according to the H3K27ac signals in HSC4 and A549 cells based on GSE143653. ( B ) Overlapping analysis of SE-associated genes in HSC4 and A549 cells. ( C ) Functional distribution of the overlapping genes.

Article Snippet: One normal human bronchial epithelial cell line (BEAS-2B) and six LUAD cell lines (H460, HCC827, A549, H1299, PC9 and Calu3) were obtained from American Type Culture Collection (ATCC, VA, USA).

Techniques: Functional Assay

Journal: International Journal of General Medicine

Article Title: Identification of Prognostic Factors Related to Super Enhancer-Regulated ceRNA Network in Metastatic Lung Adenocarcinoma

doi: 10.2147/IJGM.S332317

Figure Lengend Snippet: Effect of AC074117.1 on proliferation of LUAD cells. ( A ) qRT-PCR was used to detect the relative expression of AC074117.1 in BEAS-2B, H460, HCC827, A549, H1299, PC9 and Calu3 cells. ( B ) qRT-PCR was used to measure the knockdown efficiency of AC074117.1 in A549 and H1299 cells. ( C and D ) MTT assay was performed to evaluate changes in cell proliferation following AC074117.1 silencing in A549 and H1299 cells. **P<0.01.

Article Snippet: One normal human bronchial epithelial cell line (BEAS-2B) and six LUAD cell lines (H460, HCC827, A549, H1299, PC9 and Calu3) were obtained from American Type Culture Collection (ATCC, VA, USA).

Techniques: Quantitative RT-PCR, Expressing, Knockdown, MTT Assay

Journal: Cell chemical biology

Article Title: The Advantages of Targeted Protein Degradation over Inhibition: a RTK Case Study

doi: 10.1016/j.chembiol.2017.09.009

Figure Lengend Snippet: Key Resources Table

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies EGFR SantaCruz 1005 FLAG Sigma F1804 Tubulin Sigma T9026 HER2 Cell Signaling 2165S p-EGFR (Y1068) Abcam ab40815 p-HER2 (Y1221/1222) Cell Signaling 2243S p-AKT (T308) Cell Signaling 2965S p-ERK 1/2 (T202/204) Cell Signaling 4370 HER3 Cell Signaling 12708 pHER3 (Y1197) Cell Signaling 4561 pHER3 (Y1289) Cell Signaling 4791 pGSK-3b (S9) Cell Signaling 9331 c-Met Cell Signaling 8198 c-Met Cell Signaling 3127 p-Met (Y1245/1235) Cell Signaling 3126 p-AKT (S473) Cell Signaling 4060 Ubiquitin (P4D1) Cell Signaling 3936 p230 Trans Golgi BD Biosciences 611281 EEA1 BD Biosciences 610456 Clathrin Heavy Chain SantaCruz sc-12734 Alexa Fluor-546 conjugated anti-mouse ThermoFisher A-21143 Alexa Fluor-488 conjugated anti-rabbit ThermoFisher A-11008 HRP linked Mouse IgG GE Life Sciences NA931 HRP Linked Rabbit IgG GE Life Sciences NA934 Chemicals, Peptides, and Recombinant Proteins Cycloheximide Sigma C104450 MLN4924 Sigma 5.05477 PR-619 LifeSensors SI9619 EZ-link Sulfo-NHS-SS-Biotin Thermo 21331 Pierce NeutrAvidin Agarose beads Thermo 29200 Agarose-TUBE 1 LifeSensors UM401 Protein A-Sepharose 4B, Fast Flow beads Sigma P9424 Recombinant Human HGF Protein R&D Systems 294-HG-250 Critical Commercial Assays CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay (MTS) Promega G5421 Experimental Models: Cell Lines OVCAR8 Joyce Liu, Dana Farber MDA-MB-231 ATCC HTB-26 HeLa ATCC CCL-2 HCC827 ATCC CRL-2868 H3255 Katerina Politi, Yale H1975 ATCC CRL-5908 SKBr3 ATCC HTB-30 GTL16 F. Maina, Developmental Biology Institute of Marseille-Luminy Hs746T ATCC HTB-135 Oligonucleotides Clathrin Heavy Chain siRNA SantaCruz sc-35067 Software and Algorithms Image Lab 6.0 Biorad N/A Graphpad Prism N/A Open in a separate window Key Resources Table Contact for Reagent and Resource Sharing Further information and request for resources and reagents should be directed and will be fulfilled by Craig Crews ( ude.elay@swerc.giarc ).

Techniques: Ubiquitin Proteomics, Recombinant, Proliferation Assay, Software