title: Suppression of Aurora-A-FLJ10540 signaling axis prohibits the malignant state of head and neck cancer.
abstract: BACKGROUND
  Head and neck cancer (HNC) is a highly invasive cancer. Aurora-A has been reported for a number of malignancies. However, the identity of downstream effectors responsible for its aggressive phenotype in HNC remains underinvestigated.
  
  
  METHODS
  The mRNA and protein expression levels of Aurora-A and FLJ10540 were assessed in HNC specimens and cell lines using RT-qPCR, western blot, Oncomine, and microarray database analysis. The downstream molecular mechanisms of Aurora-A were confirmed by RT-qPCR, western blot, luciferase reporter, confocal microscopy analyses, immunoprecipitation, colony formation, cell viability, and xenograft model. Cellular functions in response to Aurora-A-modulated downstream targets such as FLJ10540 and MMPs were examined in vitro and in vivo, including cell growth, motility and chemosensitivity. Aurora-A/FLJ10540/MMPs expression was determined in cancer and adjacent normal tissues from HNC patients by immunohistochemistry approach.
  
  
  RESULTS
  In the current study, Aurora-A exhibited similar gene expression profiles with FLJ10540 by using accessibly public microarray and Oncomine database analysis, raising the possibility that these molecules might coordinately participate in cancer progression and metastasis of HNC. These two molecules connection were also examined in cell lines and tissues of HNC. Aurora-A overexpression could not only bind to the promoter of FLJ10540 to induce FLJ10540 expression, but also increase both mRNA and protein levels of MMP-7 and MMP-10 in HNC cells. Conversely, depletion of Aurora-A expression by using siRNA or Aurora-A kinase inhibitor, MLN8237, suppressed FLJ10540, MMP-7 and MMP-10 mRNA and protein expressions in vitro and in vivo. In addition, the FLJ10540-PI3K complex was destroyed by inhibition the Aurora-A kinase activity. Forced overexpression of FLJ10540 in Aurora-A-depleted or in MLN8237-treated HNC cells attenuated the effect on cytotoxicity to cisplatin. Elevated Aurora-A expression in HNC cells led to the characteristics of more aggressive malignancy, including enhanced chemoresistance and increased the abilities of proliferation, migration and invasion, which was required for FLJ10540/MMP-7 or FLJ10540/MMP-10 expressions. Finally, immunohistochemical analysis of human HNC specimens showed a significant positively correlation among Aurora-A, FLJ10540, MMP-7 and MMP-10 expressions.
  
  
  CONCLUSIONS
  Together, our findings define a novel mechanism by which Aurora-A promotes cell malignancy, with potential implications for understanding the clinical action of Aurora-A.
authors.affiliation: Center for Translational Research in Biomedical Sciences, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan. chench7@gmail.com.|Department of Otolaryngology, Kaohsiung Chang Gung Memorial Hospital, and Chang Gung University College of Medicine, Kaohsiung, Taiwan. chench7@gmail.com.|Kaohsiung Chang Gung Head and Neck Oncology Group, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan. chench7@gmail.com.|Department of Applied Chemistry and Graduate Institute of Biomedicine and Biomedical Technology, National Chi Nan University, Taoyuan, Taiwan. chench7@gmail.com.
authors.forename: Chang-Han
authors.initials: CH
authors.lastname: Chen
authors.affiliation: Institute of Physiology, National Cheng Kung University, Tainan, Taiwan. aywchang@mail.ncku.edu.tw.
authors.forename: Alice Y W
authors.initials: AY
authors.lastname: Chang
authors.affiliation: Departments of Hematology-Oncology, Chang Gung University College of Medicine, Kaohsiung, Taiwan. lee0624@cgmh.org.tw.
authors.forename: Shau-Hsuan
authors.initials: SH
authors.lastname: Li
authors.affiliation: Center for Translational Research in Biomedical Sciences, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan. thtsophia66@gmail.com.|Department of Otolaryngology, Kaohsiung Chang Gung Memorial Hospital, and Chang Gung University College of Medicine, Kaohsiung, Taiwan. thtsophia66@gmail.com.|Kaohsiung Chang Gung Head and Neck Oncology Group, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan. thtsophia66@gmail.com.
authors.forename: Hsin-Ting
authors.initials: HT
authors.lastname: Tsai
authors.affiliation: Department of Medical Research, Cell Therapy and Research Center, E-Da Hospital, I-shou University, Kaohsiung, Taiwan. her2neu24@yahoo.com.tw.
authors.forename: Li-Yen
authors.initials: LY
authors.lastname: Shiu
authors.affiliation: Graduate Institute of Systems Biology and Bioinformatics, National Central University, Jhongli, Taiwan. sulijen@gmail.com.
authors.forename: Li-Jen
authors.initials: LJ
authors.lastname: Su
authors.affiliation: Department of Otolaryngology, Kaohsiung Chang Gung Memorial Hospital, and Chang Gung University College of Medicine, Kaohsiung, Taiwan. wwenlung@gmail.com.|Kaohsiung Chang Gung Head and Neck Oncology Group, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan. wwenlung@gmail.com.|Department of Medical Research, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan. wwenlung@gmail.com.
authors.forename: Wen-Lung
authors.initials: WL
authors.lastname: Wang
authors.affiliation: Kaohsiung Chang Gung Head and Neck Oncology Group, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan. chiutaijan@gmail.com.|Departments of Hematology-Oncology, Chang Gung University College of Medicine, Kaohsiung, Taiwan. chiutaijan@gmail.com.
authors.forename: Tai-Jen
authors.initials: TJ
authors.lastname: Chiu
authors.affiliation: Department of Otolaryngology, Kaohsiung Chang Gung Memorial Hospital, and Chang Gung University College of Medicine, Kaohsiung, Taiwan. rsd0323.tw@gmail.com.|Kaohsiung Chang Gung Head and Neck Oncology Group, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan. rsd0323.tw@gmail.com.
authors.forename: Sheng-Dean
authors.initials: SD
authors.lastname: Luo
authors.affiliation: Kaohsiung Chang Gung Head and Neck Oncology Group, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan. victorhtl@yahoo.com.tw.|Departments of Hematology-Oncology, Chang Gung University College of Medicine, Kaohsiung, Taiwan. victorhtl@yahoo.com.tw.
authors.forename: Tai-Lin
authors.initials: TL
authors.lastname: Huang
authors.affiliation: Center for Translational Research in Biomedical Sciences, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan. cychien3965@cgmh.org.tw.|Department of Otolaryngology, Kaohsiung Chang Gung Memorial Hospital, and Chang Gung University College of Medicine, Kaohsiung, Taiwan. cychien3965@cgmh.org.tw.|Kaohsiung Chang Gung Head and Neck Oncology Group, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan. cychien3965@cgmh.org.tw.
authors.forename: Chih-Yen
authors.initials: CY
authors.lastname: Chien
mesh_terms: D064096:Aurora Kinase A / Q000235:genetics*; D018797:Cell Cycle Proteins / Q000235:genetics*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D002470:Cell Survival / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D006258:Head and Neck Neoplasms / Q000235:genetics*; D006801:Humans; D020782:Matrix Metalloproteinases / Q000235:genetics; D009361:Neoplasm Invasiveness / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics*; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D015398:Signal Transduction / Q000235:genetics*
chemical_list: D018797:Cell Cycle Proteins; C505378:Cep55 protein, human; D009687:Nuclear Proteins; D012333:RNA, Messenger; D019869:Phosphatidylinositol 3-Kinases; C576917:AURKA protein, human; D064096:Aurora Kinase A; D020782:Matrix Metalloproteinases