- Email: [email protected]
P3.02-084 FGF9-FGFR Pathway Induce Neuroendocrine Differentiation in Lung Epithelial Cells
S2268 together, these suggest that DKK1 induces the occurrence of EMT and promotes migration and invasion in NSCLC cells. Mechanically, b-catenin plays a vital role in DKK1-induced NSCLC cell migration and invasion, and DKK1 inhibits the phosphorylation of b-catenin, resulting in the increased nuclear localization of b-catenin.
P3.02-084 FGF9-FGFR Pathway Induce Neuroendocrine Differentiation in Lung Epithelial Cells K. Ishioka,1 H. Yasuda,2 K. Soejima,2 D. Arai,2 O. Keiko,2 J. Hamamoto,2 M. Ozaki,2 K. Masuzawa,2 I. Kawada,2 K. Naoki,2 K. Emoto,2 Y. Hayashi,3 H. Watanabe,2 H. Ahmed,2 T. Betsuyaku2 1 Pulmonary Medicine, Keio University School of Medicine, Tokyo/JP, 2 Pumonary Medicine, Keio University School of Medicine, Tokyo/JP, 3 Pathology, Keio University School of Medicine, Tokyo/JP Background: Small cell lung cancer (SCLC), an aggressive and metastatic disease, accounts for about 15% of lung cancer. Neuroendocrine differentiation is essential molecular event in SCLC development. The mechanisms of neuroendocrine differentiation and SCLC development remain elusive. For the improvement of the prognosis of SCLC patients, clarification of the mechanisms of neuroendocrine differentiation is essential. Method: For in vitro experiments, a stable cell line with constitutive expression of FGF9 in MLE12 (a mouse lung alveolar type II cell line transformed by SV40 large T antigen) was established by retroviral infections (H69: SCLC cell line, MLE12: mouse lung epithelial cell line transformed by SV40). Using these cell lines, the effect of FGF9 on proliferation, colony formation capacity and downstream signaling was evaluated by MTS assay, softagar colony formation assay and Western blotting, respectively. For in vivo experiments, these cell lines were transplanted into the immunodeficient mice subcutaneously, and the size of tumor was measured. To evaluate the efficacy of FGFR inhibitors for FGF9-driven lung cancers, AZD4547, selective FGFR inhibitor, was orally administered. For pathological characterization of the tumors, immunohistochemistry staining was performed. For patients study, 31 SCLC samples were obtained and the expression of FGF9 was evaluated by immunohistochemistry. Result: FGF9 is highly expressed in human SCLC samples (80.6%).FGF9 has oncogenic ability in vitro and its effect may be exerted by the activation of MAPK pathway through FGFR1 and FGFR3 in MLE12 cells. Unexpectedly, pathological analysis revealed FGF9-driven tumors exhibited SCLC histology. FGF9 transforms lung alveolar type II cells to SCLC in vitro and in vivo. Selective FGFR inhibitor, AZD4547 suppressed tumor growth of FGF9-driven MLE12 tumors. Conclusion: These results suggest that FGF9 has roles of tumor initiation and progression in lung cancer, especially in SCLC. SCLC which highly expresses FGF9 might be a target of FGFR inhibitors. Keywords: FGF9, FGFR, SCLC
P3.02-085 Sphingosine Kinase 1 (SPHK1) Promotes Proliferation and Survival in Non-Small Cell Lung Cancer N. Motono,1 Y. Ueda,2 A. Funasaki,1 T. Matsui,1 R. Maeda,1 A. Sekimura,1 K. Usuda,1 H. Uramoto1 1Thoracic Surgery, Kanazawa Medical University, Ishikawa/JP, 2Pathology Ii, Kanazawa Medical University, Ishikawa/JP Background: Bioactive sphingolipids, such as sphingosine-1-phosphate (S1P) and ceramide, are signaling molecules involved in the activation of pathways that are directly relevant to carcinogenesis. Sphingosine kinase (SPHK) phosphorylates sphingosine to S1P, and is therefore an important regulator of the levels of ceramide, sphingosine, and S1P. Moreover, SPHK1 has been found to up-regulated in several tumor types, such as glioblastoma multiform, intestinal adenoma, acute
Journal of Thoracic Oncology
Vol. 12 No. 11S2
erythroleukemia, prostate cancer, gastric cancer, and colon cancer. However, the relationship between bioactive sphingolipid pathway and lung cancer has not been clarified. Method: Paraffin-embedded nonsmall cell lung cancer (NSCLC) samples were obtained from 53 patients diagnosed between June 2009 and May 2014 at the Department of Thoracic surgery, Kanazawa Medical University. We characterized the SPHK1 expression in NSCLC by immunohistochemistry (IHC) and western blotting. Result: The SPHK1 expression was revealed at invasive area of NSCLC, and at fibroblast by IHC. The specific band of SPHK1 was detected approximately 52kDa by western blotting. We found that the SPHK1 expression in adenocarcinoma positively correlated with fibroblast expression and Ki-67 expression. On the other hand, the SPHK1 expression in squamous cell carcinoma was not significantly correlated with fibroblast score and Ki-67 expression. Low expression of the SPHK1 tended to prolong the relapse free survival (RFS) in lung adenocarcinoma patients (3-year RFS 100% vs 68.7%, p¼0.38). Conclusion: SPHK1 expression might regulate proliferation of cancer cell, as well as Ki-67. Furthermore, SPHK1 expression might activate fibroblast which is considered to induce epithelial mesenchymal transition. Keywords: proliferation, non-small cell lung cancer, SPHK1
P3.02-086 MGA Suppresses the MYC Pathway in Lung Adenocarcinoma P. Llabata,1 Y. Mitsuishi,2 P. Choi,2 M. Torres-Diz,1 D. Cai,2 X. Zhang,2 M. Sanchez-Cespedes,1 M. Meyerson3 1Epigenetics and Biology Program, Bellvitge Biomedical Research Institute, Barcelona/ES, 2 Medical Oncology, Dana-Farber Cancer Institute, Boston/US, 3Dana Farber Cancer Institute, Broad Institute of Harvard and Mit, and Massachusetts General Hospital, Boston, MA/US Background: Recent exome-sequencing efforts have revealed that the MGA gene, which encodes a heterodimeric partner of the MYC-interacting protein MAX, is significantly mutated (w8%) in lung adenocarcinomas. Most MGA mutations are loss-of-function, suggesting that MGA may act as a tumor suppressor. MGA mutations are mutually exclusive to MYC gene amplification, suggesting the involvement of MGA in the MYC pathway. Here, we aimed to characterize both the cellular and molecular role of MGA in lung adenocarcinoma, with a focus on studying its role in modulating the MYC pathway. Method: Chromatin immunoprecipitation-sequencing (ChIP-seq) and RNAsequencing (RNA-seq) analysis were used to identify MYC and MGA DNA binding sites and binding motifs. Immunoprecipitation assays and mass spectrometry were used to elucidate MGA gene repression mechanism. Cell competition assay was performed to measure cell
P3.02-084 FGF9-FGFR Pathway Induce Neuroendocrine Differentiation in Lung Epithelial Cells K. Ishioka,1 H. Yasuda,2 K. Soejima,2 D. Arai,2 O. Keiko,2 J. Hamamoto,2 M. Ozaki,2 K. Masuzawa,2 I. Kawada,2 K. Naoki,2 K. Emoto,2 Y. Hayashi,3 H. Watanabe,2 H. Ahmed,2 T. Betsuyaku2 1 Pulmonary Medicine, Keio University School of Medicine, Tokyo/JP, 2 Pumonary Medicine, Keio University School of Medicine, Tokyo/JP, 3 Pathology, Keio University School of Medicine, Tokyo/JP Background: Small cell lung cancer (SCLC), an aggressive and metastatic disease, accounts for about 15% of lung cancer. Neuroendocrine differentiation is essential molecular event in SCLC development. The mechanisms of neuroendocrine differentiation and SCLC development remain elusive. For the improvement of the prognosis of SCLC patients, clarification of the mechanisms of neuroendocrine differentiation is essential. Method: For in vitro experiments, a stable cell line with constitutive expression of FGF9 in MLE12 (a mouse lung alveolar type II cell line transformed by SV40 large T antigen) was established by retroviral infections (H69: SCLC cell line, MLE12: mouse lung epithelial cell line transformed by SV40). Using these cell lines, the effect of FGF9 on proliferation, colony formation capacity and downstream signaling was evaluated by MTS assay, softagar colony formation assay and Western blotting, respectively. For in vivo experiments, these cell lines were transplanted into the immunodeficient mice subcutaneously, and the size of tumor was measured. To evaluate the efficacy of FGFR inhibitors for FGF9-driven lung cancers, AZD4547, selective FGFR inhibitor, was orally administered. For pathological characterization of the tumors, immunohistochemistry staining was performed. For patients study, 31 SCLC samples were obtained and the expression of FGF9 was evaluated by immunohistochemistry. Result: FGF9 is highly expressed in human SCLC samples (80.6%).FGF9 has oncogenic ability in vitro and its effect may be exerted by the activation of MAPK pathway through FGFR1 and FGFR3 in MLE12 cells. Unexpectedly, pathological analysis revealed FGF9-driven tumors exhibited SCLC histology. FGF9 transforms lung alveolar type II cells to SCLC in vitro and in vivo. Selective FGFR inhibitor, AZD4547 suppressed tumor growth of FGF9-driven MLE12 tumors. Conclusion: These results suggest that FGF9 has roles of tumor initiation and progression in lung cancer, especially in SCLC. SCLC which highly expresses FGF9 might be a target of FGFR inhibitors. Keywords: FGF9, FGFR, SCLC
P3.02-085 Sphingosine Kinase 1 (SPHK1) Promotes Proliferation and Survival in Non-Small Cell Lung Cancer N. Motono,1 Y. Ueda,2 A. Funasaki,1 T. Matsui,1 R. Maeda,1 A. Sekimura,1 K. Usuda,1 H. Uramoto1 1Thoracic Surgery, Kanazawa Medical University, Ishikawa/JP, 2Pathology Ii, Kanazawa Medical University, Ishikawa/JP Background: Bioactive sphingolipids, such as sphingosine-1-phosphate (S1P) and ceramide, are signaling molecules involved in the activation of pathways that are directly relevant to carcinogenesis. Sphingosine kinase (SPHK) phosphorylates sphingosine to S1P, and is therefore an important regulator of the levels of ceramide, sphingosine, and S1P. Moreover, SPHK1 has been found to up-regulated in several tumor types, such as glioblastoma multiform, intestinal adenoma, acute
Journal of Thoracic Oncology
Vol. 12 No. 11S2
erythroleukemia, prostate cancer, gastric cancer, and colon cancer. However, the relationship between bioactive sphingolipid pathway and lung cancer has not been clarified. Method: Paraffin-embedded nonsmall cell lung cancer (NSCLC) samples were obtained from 53 patients diagnosed between June 2009 and May 2014 at the Department of Thoracic surgery, Kanazawa Medical University. We characterized the SPHK1 expression in NSCLC by immunohistochemistry (IHC) and western blotting. Result: The SPHK1 expression was revealed at invasive area of NSCLC, and at fibroblast by IHC. The specific band of SPHK1 was detected approximately 52kDa by western blotting. We found that the SPHK1 expression in adenocarcinoma positively correlated with fibroblast expression and Ki-67 expression. On the other hand, the SPHK1 expression in squamous cell carcinoma was not significantly correlated with fibroblast score and Ki-67 expression. Low expression of the SPHK1 tended to prolong the relapse free survival (RFS) in lung adenocarcinoma patients (3-year RFS 100% vs 68.7%, p¼0.38). Conclusion: SPHK1 expression might regulate proliferation of cancer cell, as well as Ki-67. Furthermore, SPHK1 expression might activate fibroblast which is considered to induce epithelial mesenchymal transition. Keywords: proliferation, non-small cell lung cancer, SPHK1
P3.02-086 MGA Suppresses the MYC Pathway in Lung Adenocarcinoma P. Llabata,1 Y. Mitsuishi,2 P. Choi,2 M. Torres-Diz,1 D. Cai,2 X. Zhang,2 M. Sanchez-Cespedes,1 M. Meyerson3 1Epigenetics and Biology Program, Bellvitge Biomedical Research Institute, Barcelona/ES, 2 Medical Oncology, Dana-Farber Cancer Institute, Boston/US, 3Dana Farber Cancer Institute, Broad Institute of Harvard and Mit, and Massachusetts General Hospital, Boston, MA/US Background: Recent exome-sequencing efforts have revealed that the MGA gene, which encodes a heterodimeric partner of the MYC-interacting protein MAX, is significantly mutated (w8%) in lung adenocarcinomas. Most MGA mutations are loss-of-function, suggesting that MGA may act as a tumor suppressor. MGA mutations are mutually exclusive to MYC gene amplification, suggesting the involvement of MGA in the MYC pathway. Here, we aimed to characterize both the cellular and molecular role of MGA in lung adenocarcinoma, with a focus on studying its role in modulating the MYC pathway. Method: Chromatin immunoprecipitation-sequencing (ChIP-seq) and RNAsequencing (RNA-seq) analysis were used to identify MYC and MGA DNA binding sites and binding motifs. Immunoprecipitation assays and mass spectrometry were used to elucidate MGA gene repression mechanism. Cell competition assay was performed to measure cell