MET Increases the Sensitivity of Gefitinib-Resistant Cells to SN-38, an Active Metabolite of Irinotecan, by Up-Regulating the Topoisomerase I Activity

Slides:

Advertisements

Similar presentations

Nicotinamide Phosphoribosyltransferase: A Potent Therapeutic Target in Non-small Cell Lung Cancer with Epidermal Growth Factor Receptor-Gene Mutation

Advertisements

Inducible EGFR T790M-Mediated Gefitinib Resistance in Non-small Cell Lung Cancer Cells Does Not Modulate Sensitivity to PI103 Provoked Autophagy Flavia.

IL-18 Downregulates Collagen Production in Human Dermal Fibroblasts via the ERK Pathway Hee Jung Kim, Seok Bean Song, Jung Min Choi, Kyung Moon Kim,

Molecular Therapy - Oncolytics

The Autophagy Inhibitor Chloroquine Overcomes the Innate Resistance of Wild-Type EGFR Non-Small-Cell Lung Cancer Cells to Erlotinib Yiyu Zou, PhD, Yi-He.

The Epidermal Growth Factor Receptor-L861Q Mutation Increases Kinase Activity without Leading to Enhanced Sensitivity Toward Epidermal Growth Factor Receptor.

NDRG1/Cap43/Drg-1 may Predict Tumor Angiogenesis and Poor Outcome in Patients with Lung Cancer Koichi Azuma, MD, PhD, Akihiko Kawahara, PhD, Satoshi.

Group IIa secretory phospholipase expression correlates with group IIa secretory phospholipase inhibition–mediated cell death in K-ras mutant lung cancer.

Upregulation of PD-L1 by EGFR Activation Mediates the Immune Escape in EGFR- Driven NSCLC: Implication for Optional Immune Targeted Therapy for NSCLC Patients.

Volume 133, Issue 2, Pages (August 2007)

Nicotine Induces Resistance to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor by α1 Nicotinic Acetylcholine Receptor–Mediated Activation in.

Roles of the cytoplasmic domains of the α and β subunits of human granulocyte- macrophage colony-stimulating factor receptor Akihiko Muto, PhDa, Sumiko.

Rafal Dziadziuszko, MD, PhD, Anh T

Volume 128, Issue 1, Pages (January 2005)

Resistance to EGFR-TKI Can Be Mediated through Multiple Signaling Pathways Converging upon Cap-Dependent Translation in EGFR-Wild Type NSCLC Manish R.

Constitutive and tumor necrosis factor-α-induced activation of nuclear factor-κB in adenomyosis and its inhibition by andrographolide Bin Li, M.S., Ming.

Keloid Fibroblasts Resist Ceramide-Induced Apoptosis by Overexpression of Insulin- Like Growth Factor I Receptor Hiroshi Ishihara, Hiroshi Yoshimoto,

Volume 69, Issue 4, Pages (February 2006)

Complement Factor H: A Biomarker for Progression of Cutaneous Squamous Cell Carcinoma Pilvi M. Riihilä, Liisa M. Nissinen, Risto Ala-aho, Markku Kallajoki,

Interleukin-17 and Prostaglandin E2 Are Involved in Formation of an M2 Macrophage- Dominant Microenvironment in Lung Cancer Lunxu Liu, MD, PhD, Dongxia.

Characterization of Fibroblast Growth Factor Receptor 1 in Small-Cell Lung Cancer Anish Thomas, MD, Jih-Hsiang Lee, MD, Zied Abdullaev, PhD, Kang-Seo.

Yu-Hsin Chiu, Jennifer Y. Lee, Lewis C. Cantley Molecular Cell

MET Increases the Sensitivity of Gefitinib-Resistant Cells to SN-38, an Active Metabolite of Irinotecan, by Up-Regulating the Topoisomerase I Activity

Activated Leukocyte Cell-Adhesion Molecule (ALCAM) Promotes Malignant Phenotypes of Malignant Mesothelioma Futoshi Ishiguro, MD, Hideki Murakami, MD,

The C-terminus of Hsp70-Interacting Protein Promotes Met Receptor Degradation Kang Won Jang, PhD, Jeong Eun Lee, MD, Sun Young Kim, MD, Min-Woong Kang,

Inducible EGFR T790M-Mediated Gefitinib Resistance in Non-small Cell Lung Cancer Cells Does Not Modulate Sensitivity to PI103 Provoked Autophagy Flavia.

Combining the Multitargeted Tyrosine Kinase Inhibitor Vandetanib with the Antiestrogen Fulvestrant Enhances Its Antitumor Effect in Non-small Cell Lung.

Frequent Coamplification and Cooperation between C-MYC and PVT1 Oncogenes Promote Malignant Pleural Mesothelioma Erick Riquelme, PhD, Milind B. Suraokar,

The Autophagy Inhibitor Chloroquine Overcomes the Innate Resistance of Wild-Type EGFR Non-Small-Cell Lung Cancer Cells to Erlotinib Yiyu Zou, PhD, Yi-He.

Hepatocyte Growth Factor Induces Resistance to Anti-Epidermal Growth Factor Receptor Antibody in Lung Cancer Tadaaki Yamada, MD, PhD, Shinji Takeuchi,

Nicotine Induces Resistance to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor by α1 Nicotinic Acetylcholine Receptor–Mediated Activation in.

Ha Linh Vu, Sheera Rosenbaum, Claudia Capparelli, Timothy J

Glucocorticoid Receptor Status Is a Principal Determinant of Variability in the Sensitivity of Non–Small-Cell Lung Cancer Cells to Pemetrexed Mugdha.

Stefan W. Stoll, Jessica L. Johnson, Yong Li, Laure Rittié, James T

Expression and regulation of Toll-like receptor 2 by IL-1β and fibronectin fragments in human articular chondrocytes S.-L. Su, M.S., C.-D. Tsai, Ph.D.,

Hsp90 Inhibition Overcomes HGF-Triggering Resistance to EGFR-TKIs in EGFR-Mutant Lung Cancer by Decreasing Client Protein Expression and Angiogenesis

Akio Horiguchi, Mototsugu Oya, Ken Marumo, Masaru Murai

Interleukin-22 Is Frequently Expressed in Small- and Large-Cell Lung Cancer and Promotes Growth in Chemotherapy-Resistant Cancer Cells Sebastian Kobold,

MicroRNA-101 Exerts Tumor-Suppressive Functions in Non-small Cell Lung Cancer through Directly Targeting Enhancer of Zeste Homolog 2 Ji-guang Zhang,

Lovastatin Sensitizes Lung Cancer Cells to Ionizing Radiation: Modulation of Molecular Pathways of Radioresistance and Tumor Suppression Toran Sanli,

The Epidermal Growth Factor Receptor-L861Q Mutation Increases Kinase Activity without Leading to Enhanced Sensitivity Toward Epidermal Growth Factor Receptor.

Downregulated ABCG2 Enhances Sensitivity to Topoisomerase I Inhibitor in Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor-Resistant Non-small.

Volume 19, Issue 1, Pages (January 2011)

Plexin B1 Suppresses c-Met in Melanoma: A Role for Plexin B1 as a Tumor-Suppressor Protein through Regulation of c-Met Laurel Stevens, Lindy McClelland,

Sema4D, the Ligand for Plexin B1, Suppresses c-Met Activation and Migration and Promotes Melanocyte Survival and Growth Joanne Soong, Yulin Chen, Elina.

IL-6 Secreted from Cancer-Associated Fibroblasts Mediates Chemoresistance in NSCLC by Increasing Epithelial-Mesenchymal Transition Signaling Yasushi.

1,25-dihydroxyvitamin D3 inhibits renal interstitial myofibroblast activation by inducing hepatocyte growth factor expression Yingjian Li, Bradley C.

Volume 132, Issue 5, Pages (May 2007)

Profiling Motility Signal-Specific Genes in Primary Human Keratinocytes Chieh-Fang Cheng, Jianhua Fan, Balaji Bandyopahdhay, Dennis Mock, Shengxi Guan,

Erratum Journal of Thoracic Oncology

Post-Transcriptional Regulation of Melanin Biosynthetic Enzymes by cAMP and Resveratrol in Human Melanocytes Richard A. Newton, Anthony L. Cook, Donald.

Triple Inhibition of EGFR, Met, and VEGF Suppresses Regrowth of HGF-Triggered, Erlotinib-Resistant Lung Cancer Harboring an EGFR Mutation Junya Nakade,

Nicotinamide Phosphoribosyltransferase: A Potent Therapeutic Target in Non-small Cell Lung Cancer with Epidermal Growth Factor Receptor-Gene Mutation

Volume 39, Issue 3, Pages (September 2003)

Coactivation of Receptor Tyrosine Kinases in Malignant Mesothelioma as a Rationale for Combination Targeted Therapy Marie Brevet, MD, PhD, Shigeki Shimizu,

Roles of the cytoplasmic domains of the α and β subunits of human granulocyte- macrophage colony-stimulating factor receptor Akihiko Muto, PhDa, Sumiko.

IL-18 Downregulates Collagen Production in Human Dermal Fibroblasts via the ERK Pathway Hee Jung Kim, Seok Bean Song, Jung Min Choi, Kyung Moon Kim,

Collagen Synthesis Is Suppressed in Dermal Fibroblasts by the Human Antimicrobial Peptide LL-37 Hyun Jeong Park, Dae Ho Cho, Hee Jung Kim, Jun Young.

IL-12 affects Dermatophagoides farinae–induced IL-4 production by T cells from pediatric patients with mite-sensitive asthma Takeshi Noma, MD, PhD, Izumi.

Volume 67, Issue 6, Pages (June 2005)

Epithelial to Mesenchymal Transition in an Epidermal Growth Factor Receptor-Mutant Lung Cancer Cell Line with Acquired Resistance to Erlotinib Kenichi.

Volume 56, Issue 5, Pages (November 1999)

Volume 18, Issue 3, Pages (March 2010)

Intestinal myofibroblasts in innate immune responses of the intestine

Histone deacetylase inhibitors down-regulate G-protein-coupled estrogen receptor and the GPER-antagonist G-15 inhibits proliferation in endometriotic.

Volume 55, Issue 2, Pages (February 1999)

Bcl-2 and bcl-xL Antisense Oligonucleotides Induce Apoptosis in Melanoma Cells of Different Clinical Stages Robert A. Olie, Christoph Hafner, Renzo Küttel,

Molecular Therapy - Oncolytics

Signal Transducer and Activator of Transcription 3 as Molecular Therapy for Non–Small- Cell Lung Cancer Cheng-Yi Wang, MD, Ting-Ting Chao, PhD, Wei-Tien.

Expression and induction of HER2 and HPSE in 231BMBC cells.

Presentation transcript:

MET Increases the Sensitivity of Gefitinib-Resistant Cells to SN-38, an Active Metabolite of Irinotecan, by Up-Regulating the Topoisomerase I Activity Asao Sakai, MD, Kazuo Kasahara, PhD, Tohru Ohmori, PhD, Hideharu Kimura, PhD, Takashi Sone, PhD, Masaki Fujimura, PhD, Shinji Nakao, PhD Journal of Thoracic Oncology Volume 7, Issue 9, Pages 1337-1344 (September 2012) DOI: 10.1097/JTO.0b013e31825cca4c Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

Figure 1 The sensitivity of PC-9 and gefitinib-resistant PC-9/Met cells to gefitinib and SN-38. The cells (2 × 103 cells/well) were seeded on 96-well plates, and after preincubation for 24 hours were exposed to various concentrations of each drug. After incubation for 72 hours, the growth inhibition rate was analyzed by a WST-1 assay as described in the Materials and Methods section. The points represent the means of the data generated from at least three experiments performed in triplicate. Bars, SD. PC-9 cells, PC-9/Met cells. SN-38, 7-ethyl-10-hydroxy-camptothecin. Journal of Thoracic Oncology 2012 7, 1337-1344DOI: (10.1097/JTO.0b013e31825cca4c) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

Figure 2 A, A comparison of the protein expression levels of topo I, MET, p-MET, EGFR, p-EGFR, Akt, and p-Akt in the PC-9 and PC-9/Met cells as determined by a Western blot analysis. Topo I migrated to approximately the 70 kDa position. The data are expressed as the means ± SD from more than three experiments. *p < 0.05. B. The MET gene amplification status in the PC-9 and PC-9/Met cells was determined by a fluorescence in situ hybridization analysis. The chromosome 7 centromere signals are green, and the MET signals are red. MET amplification was found in the PC-9/Met cells, but not in the PC-9 cells. C, Topo I activity in the PC-9 cells, and the gefitinib-resistant PC-9/Met cells. Complete activity is seen when no supercoiled DNA substrate remains, and the presence of some supercoiled DNA indicates partial activity. The upper bands are attributable to DNA nicking by contaminating nucleases. The bottom band, indicated by the arrow, represents supercoiled DNA degraded by topo I. The topo I activity was normalized to the amount of nuclear protein extracted, such as 1.8 μg, 0.9 μg, 0.45 μg, and 0.225 μg, respectively, in both cell lines. In the PC-9/Met cells, the band of supercoiled DNA was eliminated when 0.9 μg of nuclear protein was added to the reaction mixture. However, the band of supercoiled DNA remained when 1.8 μg of nuclear protein from the PC-9 cells was added. These results showed that the topo I activity of the PC-9/Met cells was approximately fourfold higher than that of the PC-9 cells. D, Measurement of the topo I mRNA expression in PC-9 and PC-9/Met cells as determined by real-time PCR. The topo I mRNA level of the PC-9/Met cells was approximately 9.2 times higher than that of the PC-9 cells. EGFR, epidermal growth receptor factor; p-EFGR, phospho-EGFR; p-Akt, phospho-Akt; p-MET, phospho-MET; topo I, topoisomerase I; mRNA, messenger RNA; PCR, polymerase chain reaction. Journal of Thoracic Oncology 2012 7, 1337-1344DOI: (10.1097/JTO.0b013e31825cca4c) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

Figure 3 Specific down-regulation of MET affects the level of topo I protein expression and the enzyme activity, while also decreasing the sensitivity of cells to SN-38. Control and MET-specific siRNA were introduced into the PC-9 and PC-9/Met cell lines. A. Whole-cell proteins and nuclear proteins were extracted 48 hours later, and a Western blot analysis was performed. To detect the MET protein, a 20-μg protein sample was loaded per lane for the PC-9 cells and PC-9/Met cells. To measure topo I protein expression, 10-μg samples of nuclear protein were loaded for both the PC-9 and PC-9/Met cell lines. The data are expressed as the means ± SD from more than three experiments, *p < 0.05 versus control. B. The relationship between topo I expression and MET expression by a Western blotting analysis at 0 hour, 6 hours, 12 hours, and 24 hours after siRNA addition. C, The effect of the specific down-regulation of MET on the topo I activity of the PC-9/Met cells. MET-specific siRNA was introduced into PC-9/Met cells, and nuclear enzymes were extracted from the PC-9/Met cells that contained topo I. The PC-9/Met cells in which MET had been down-regulated had lower topo I activity than did the untreated PC-9/Met cells. D, The results of the WST-1 assay of cell-growth inhibition by SN-38. The PC-9/Met cells were more sensitive to SN-38 than the PC-9 cells, and the sensitivity of cells treated with MET siRNA was reduced. The points represent the means of data generated from at least three experiments performed in triplicate. Bars, SD. PC-9, PC-9/Met, PC-9/Met siRNA. topo I, topoisomerase I; siRNA, small interfering RNA, SN-38, 7-ethyl-10-hydroxy-camptothecin; con, control; scr, scramble. Journal of Thoracic Oncology 2012 7, 1337-1344DOI: (10.1097/JTO.0b013e31825cca4c) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

Figure 4 A, Cell extracts from PC-9 cells and PC-9/Met cells were prepared after the cells were cultured with HGF (20 ng/ml) for 30 minutes and 60 minutes, after serum starvation for 24 hours. HGF stimulation induced MET phosphorylation in both cell lines. Nuclear extracts containing the topo I enzyme were prepared after 6 hours, 12 hours, and 24 hours of HGF stimulation, and the topo I protein expression level in both PC-9 and PC-9/Met cells was increased after HGF stimulation. The data are expressed as the means ± SD from three experiments, *p < 0.05 versus control. B, HGF increased the sensitivity of PC-9 and PC-9/Met cells to SN-38. Tumor cells at concentration of 2 × 104 cells/well were incubated in RPMI medium supplemented with 0.1% fetal bovine serum overnight. HGF (20 ng/ml) was added to the cultures of tumor cells for 4 hours. Increasing concentrations of SN-38 were added to the each well, and the incubation was continued for a further 24 hours. Then, the cell growth was determined by the water soluble tetrazolium salt-1 assay. HGF, hepatocyte growth factor; topo I, topoisomerase 1. Journal of Thoracic Oncology 2012 7, 1337-1344DOI: (10.1097/JTO.0b013e31825cca4c) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions