VEGFA Upregulates FLJ10540 and Modulates Migration and Invasion of Lung Cancer via PI3K/AKT Pathway Chang-Han Chen1, Jin-Mei Lai2, Teh-Ying Chou 3 , Cheng-Yu Chen3, Li-Jen Su 4 , Yuan-Chii Lee5, Tai-Shan Cheng 6 , Yi-Ren Hong 6 , Chen-Kung Chou7, Jacqueline Whang-Peng8*, Yu-Chung Wu4*, Chi-Ying F. 3 Huang * 1 Department of Otolaryngology and Kaohsiung Chang Gung Head and Neck Oncology Group, Chang Gung Memorial Hospital-Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung, Taiwan, 2 Department of Life Science, Fu-Jen Catholic University, Taipei Hsien, Taiwan, 3 Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan, 4 Division of Thoracic Surgery, Department of Surgery, Veterans General Hospital, Taipei, Taiwan, 5 Graduate Institute of Medical Informatics, Taipei Medical University, Taipei, Taiwan, 6 Graduate Institute of Biochemistry, Kaohsiung Medical University, Kaohsiung, Taiwan, 7 Department of Life Science, Chang Gung University, Tao-Yuan, Taiwan, 8 Division of Cancer Center, Wan Fang Hospital, Taipei, Taiwan Abstract Background: Lung adenocarcinoma is the leading cause of cancer-related deaths among both men and women in the world. Despite recent advances in diagnosis and treatment, the mortality rates with an overall 5-year survival of only 15%. This high mortality is probably attributable to early metastasis. Although several well-known markers correlated with poor/ metastasis prognosis in lung adenocarcinoma patients by immunohistochemistry was reported, the molecular mechanisms of lung adenocarcinoma development are still not clear. To explore novel molecular markers and their signaling pathways will be crucial for aiding in treatment of lung adenocarcinoma patients. Methodology/Principal Findings: To identify novel lung adenocarcinoma-associated /metastasis genes and to clarify the underlying molecular mechanisms of these targets in lung cancer progression, we created a bioinformatics scheme consisting of integrating three gene expression profile datasets, including pairwise lung adenocarcinoma, secondary metastatic tumors vs. benign tumors, and a series of invasive cell lines. Among the novel targets identified, FLJ10540 was overexpressed in lung cancer tissues and is associated with cell migration and invasion. Furthermore, we employed two co- expression strategies to identify in which pathway FLJ10540 was involved. Lung adenocarcinoma array profiles and tissue microarray IHC staining data showed that FLJ10540 and VEGF-A, as well as FLJ10540 and phospho-AKT exhibit positive correlations, respectively. Stimulation of lung cancer cells with VEGF-A results in an increase in FLJ10540 protein expression and enhances complex formation with PI3K. Treatment with VEGFR2 and PI3K inhibitors affects cell migration and invasion by activating the PI3K/AKT pathway. Moreover, knockdown of FLJ10540 destabilizes formation of the P110-a/P85-a-(PI3K) complex, further supporting the participation of FLJ10540 in the VEGF-A/PI3K/AKT pathway. Conclusions/Significance: This finding set the stage for further testing of FLJ10540 as a new therapeutic target for treating lung cancer and may contribute to the development of new therapeutic strategies that are able to block the PI3K/AKT pathway in lung cancer cells. Citation: Chen C-H, Lai J-M, Chou T-Y, Chen C-Y, Su L-J, et al. (2009) VEGFA Upregulates FLJ10540 and Modulates Migration and Invasion of Lung Cancer via PI3K/ AKT Pathway. PLoS ONE 4(4): e5052. doi:10.1371/journal.pone.0005052 Editor: Mikhail V. Blagosklonny, Ordway Research Institute, United States of America Received December 22, 2008; Accepted February 12, 2009; Published April 1, 2009 Copyright: 2009 Chen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This study was supported in part by grants from the Chang Gung Hospital (CMRPD140211) and NSC (94-2752-B-010-002-PAE) to Chen-Kung Chou, NSC (Program for Interdisciplinary Research Project: NSC97-2627-B-010-011 to Chi-Ying Huang, NSC97-2627-B-030-001 to J.-M. Lai), and NSC97-3112-B-010-025-CC1, Taichung Veteran General Hospital (TCVGH-957326D), and the Program for Promoting Academic Excellence of Universities (National Yang Ming University) to Chi- Ying Huang, and NSC (95-3112-B-075-002 and 96-3112-B-075-001) to Yu-Chung Wu. The Gene Expression Analysis Core Facility is supported by the National Research Program for Genomic Medicine (NRPGM), National Science Council, Taiwan. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: jqwpeng@tmu.edu.tw (JW-P); wuyc@vghtpe.gov.tw (Y-CW); cyhuang5@ym.edu.tw (C-YFH) Introduction Lung cancer is the leading cause of cancer-related deaths among both men and women in the world [1–2]. Despite recent advances in diagnosis and treatment, the mortality rates remain high, with an overall 5-year survival of only 15%. Surgery remains the first choice of treatment for localized non-small cell lung cancer and provides the best opportunity for cure. However, when first diagnosed, most patients already have advanced disease, and only 35% of patients with non-small cell lung cancer (NSCLC) are eligible for resection [3]. Novel molecular markers or targets aiding in diagnosis and treatment will be crucial for improving the mortality rate. Tumor invasion and metastasis are important areas for study in order to determine the aggressive phenotype of human cancers and are the major causes of cancer deaths [4]. The process of metastasis is very complex and is considered a late event in tumorigenesis, i.e. cells proliferate, lose contact with neighboring PLoS ONE | www.plosone.org 1 April 2009 | Volume 4 | Issue 4 | e5052