title: Inhibitory effect oftrans-ferulic acid on proliferation and migration of human lung cancer cells accompanied with increased endogenous reactive oxygen species and β-catenin instability.
abstract: BACKGROUND
  Trans-ferulic (FA) acid exhibits antioxidant effects in vitro. However, the underlying mechanism oftrans-FA activity in cellular physiology, especially cancer physiology, remains largely unknown. This study investigated the cellular physiological effects oftrans-FA on the H1299 human lung cancer cell line.
  
  
  METHODS
  The 2,2-diphenyl-1-picrylhydrazyl assay was used to determine free radical scavenging capability. Assessment of intracellular reactive oxygen species (ROS) was evaluated using oxidized 2',7'-dichlorofluorescin diacetate and dihydroethidium staining. Trypan blue exclusion, colony formation, and anchorage-independent growth assays were used to determine cellular proliferation. Annexin V staining assay was used to assess cellular apoptosis by flow cytometry. Wound healing and Boyden's well assays were used to detect the migration and invasion of cells. Gelatin zymography was used to detect matrix metalloproteinase (MMP-2 and MMP-9) activity. Western blotting was used to detect expression levels of various signaling pathway proteins.
  
  
  RESULTS
  DPPH assay results indicated thattrans-FA exerted potent antioxidant effects. However,trans-FA increased intracellular ROS levels, including hydrogen peroxide and superoxide anion, in H1299 cells.Trans-FA treatment inhibited cellular proliferation and induced moderate apoptotic cell death at the highest concentration used (0.6 mM). Furthermore,trans-FA moderately inhibited the migration of H1299 cells at the concentrations of 0.3 and 0.6 mM and attenuated MMP-2 and MMP-9 activity.Trans-FA caused the phosphorylation of β-catenin, resulting in proteasomal degradation of β-catenin. Conversely,trans-FA treatment increased the expression of pro-apoptotic factor Bax and decreased the expression of pro-survival factor survivin.
  
  
  CONCLUSIONS
  Various concentrations (0.06-0.6 mM) oftrans-FA exert both anti-proliferation and anti-migration effects in the human lung cancer cell line H1299.
authors.affiliation: Department of Thoracic Surgery, Chi-Mei Medical Center, Tainan, 710 Taiwan.
authors.forename: Yao
authors.initials: Y
authors.lastname: Fong
authors.affiliation: Division of Chest, Ten Chan General Hospital, Chung-Li, 320 Taiwan, ROC.
authors.forename: Chia-Chun
authors.initials: CC
authors.lastname: Tang
authors.affiliation: Department of Biotechnology, Kaohsiung Medical University, Kaohsiung, 807 Taiwan.
authors.forename: Huei-Ting
authors.initials: HT
authors.lastname: Hu
authors.affiliation: Department of Food Nutrition, Chung-Hwa University of Medical Technology, Tainan, 701 Taiwan.
authors.forename: Hsin-Yu
authors.initials: HY
authors.lastname: Fang
authors.affiliation: Department of Biotechnology, Kaohsiung Medical University, Kaohsiung, 807 Taiwan.|The Institute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung, 804 Taiwan.
authors.forename: Bing-Hung
authors.initials: BH
authors.lastname: Chen
authors.affiliation: Department of Biotechnology, Kaohsiung Medical University, Kaohsiung, 807 Taiwan.|Department of Biological Sciences, National Sun Yat-sen University, Kaohsiung, 804 Taiwan.
authors.forename: Chang-Yi
authors.initials: CY
authors.lastname: Wu
authors.affiliation: Translational Research Center, Kaohsiung Medical University Hospital, Kaohsiung, 807 Taiwan.
authors.forename: Shyng-Shiou
authors.initials: SS
authors.lastname: Yuan
authors.affiliation: Graduate Institute of Biomedical Engineering, National Chung Hsing University, Taichung, 402 Taiwan.
authors.forename: Hui-Min David
authors.initials: HD
authors.lastname: Wang
authors.affiliation: Department of Biotechnology, Kaohsiung Medical University, Kaohsiung, 807 Taiwan.
authors.forename: Yen-Chun
authors.initials: YC
authors.lastname: Chen
authors.affiliation: Department of Biological Sciences and Technology, National University of Tainan, Tainan, 700 Taiwan.
authors.forename: Yen-Ni
authors.initials: YN
authors.lastname: Teng
authors.affiliation: Department of Biotechnology, Kaohsiung Medical University, Kaohsiung, 807 Taiwan.|Department of Biological Sciences, National Sun Yat-sen University, Kaohsiung, 804 Taiwan.|Translational Research Center, Kaohsiung Medical University Hospital, Kaohsiung, 807 Taiwan.|Research Center for Environment Medicine, Kaohsiung Medical University, Kaohsiung, 807 Taiwan.
authors.forename: Chien-Chih
authors.initials: CC
authors.lastname: Chiu