title: A proteomics approach to identifying novel protein targets involved in erinacine A-mediated inhibition of colorectal cancer cells' aggressiveness.
abstract: Erinacine A, a major active component of a diterpenoid derivative isolated from Hericium erinaceus mycelium, has been demonstrated to exert anticancer effects. Herein, we present an investigation of the molecular mechanism of erinacine A induction associated with cancer cells' aggressive status and death. A proteomic approach was used to purify and identify the differentially expressed proteins following erinacine A treatment and the mechanism of its action in apoptotic and the targets of erinacine A. Our results demonstrate that erinacine A treatment of HCT-116 and DLD-1 cells increased cell cytotoxicity and reactive oxygen species (ROS) production as well as decreased cell proliferation and invasiveness. Ten differentially displayed proteins were determined and validated in vitro and in vivo between the erinacine A-treated and untreated groups. In addition, erinacine A time-dependent induction of cell death and inhibitory invasiveness was associated with sustained phosphorylation of the PI3K/mTOR/p70S6K and ROCK1/LIMK2/Cofilin pathways. Furthermore, we demonstrated that erinacine A-induced HCT-116 and DLD-1 cells viability and anti-invasion properties by up-regulating the activation of PI3K/mTOR/p70S6K and production of ROS. Experiments involving specific inhibitors demonstrated that the differential expression of cofilin-1 (COFL1) and profilin-1 (PROF1) during erinacine A treatment could be involved in the mechanisms of HCT-116 and DLD-1 cells death and decreased aggressiveness, which occurred via ROCK1/LIMK2/Cofilin expression, with activation of the PI3K/mTOR/p70S6K signalling pathway. These findings elucidate the mechanism of erinacine A inhibiting the aggressive status of cells by activating PI3K/mTOR/p70S6K downstream signalling and the novel protein targets COF1 and PROF1; this could be a good molecular strategy to limit the aggressiveness of CRC cells.
authors.affiliation: Division of Colorectal Surgery, Department of Surgery, Chang Gung Memorial Hospital, Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung, Taiwan.
authors.forename: Ko-Chao
authors.initials: KC
authors.lastname: Lee
authors.affiliation: Department of Nursing, Chang Gung University of Science and Technology, Chiayi, Taiwan.|Research Center for Industry of Human Ecology, Chang Gung University of Science and Technology, Taoyuan, Taiwan.|Chronic Diseases and Health Promotion Research Center, CGUST, Chiayi, Taiwan.
authors.forename: Hsing-Chun
authors.identifier: 0000-0002-8993-1615
authors.initials: HC
authors.lastname: Kuo
authors.affiliation: Department of Hepato-Gastroenterology, Chang Gung Memorial Hospital, Chiayi, Taiwan.|Graduate Institute of Clinical Medical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Chien-Heng
authors.initials: CH
authors.lastname: Shen
authors.affiliation: Division of Colorectal Surgery, Department of Surgery, Chang Gung Memorial Hospital, Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung, Taiwan.
authors.forename: Chien-Chang
authors.initials: CC
authors.lastname: Lu
authors.affiliation: Division of Colon and Rectal Surgery, Department of Surgery, Chang Gung Memorial Hospital Chiayi, Chiayi, Taiwan.|Chang Gung University College of Medicine, Taoyuan, Taiwan.
authors.forename: Wen-Shih
authors.initials: WS
authors.lastname: Huang
authors.affiliation: Graduate Institute of Clinical Medical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.|Division of Colon and Rectal Surgery, Department of Surgery, Chang Gung Memorial Hospital Chiayi, Chiayi, Taiwan.
authors.forename: Meng-Chiao
authors.initials: MC
authors.lastname: Hsieh
authors.affiliation: Division of Colon and Rectal Surgery, Department of Surgery, Chang Gung Memorial Hospital Chiayi, Chiayi, Taiwan.
authors.forename: Cheng-Yi
authors.initials: CY
authors.lastname: Huang
authors.affiliation: Division of Colon and Rectal Surgery, Department of Surgery, Chang Gung Memorial Hospital Chiayi, Chiayi, Taiwan.
authors.forename: Yi-Hung
authors.initials: YH
authors.lastname: Kuo
authors.affiliation: Department of Hepato-Gastroenterology, Chang Gung Memorial Hospital, Chiayi, Taiwan.
authors.forename: Yung-Yu
authors.initials: YY
authors.lastname: Hsieh
authors.affiliation: Department of Nursing, Chang Gung University of Science and Technology, Chiayi, Taiwan.
authors.forename: Chih-Chuan
authors.initials: CC
authors.lastname: Teng
authors.affiliation: Grape King Biotechnology Inc (Grape King Bio Ltd.), Zhong-Li, Taiwan.
authors.forename: Li-Ya
authors.initials: LY
authors.lastname: Lee
authors.affiliation: Department of Hepato-Gastroenterology, Chang Gung Memorial Hospital, Chiayi, Taiwan.|Chang Gung University College of Medicine, Taoyuan, Taiwan.
authors.forename: Shui-Yi
authors.initials: SY
authors.lastname: Tung
mesh_terms: D051339:Actin Depolymerizing Factors / Q000378:metabolism; D000818:Animals; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D015179:Colorectal Neoplasms / Q000378:metabolism*; D004224:Diterpenes / Q000494:pharmacology*; D005260:Female; D045325:HCT116 Cells; D006801:Humans; D054461:Lim Kinases / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009361:Neoplasm Invasiveness / Q000473:pathology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D051304:Profilins / Q000378:metabolism; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D017382:Reactive Oxygen Species / Q000378:metabolism; D038762:Ribosomal Protein S6 Kinases, 70-kDa / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism; D054460:rho-Associated Kinases / Q000378:metabolism
chemical_list: D051339:Actin Depolymerizing Factors; D004224:Diterpenes; D054461:Lim Kinases; D019869:Phosphatidylinositol 3-Kinases; D051304:Profilins; D020543:Proteome; D017382:Reactive Oxygen Species; D038762:Ribosomal Protein S6 Kinases, 70-kDa; D058570:TOR Serine-Threonine Kinases; D054460:rho-Associated Kinases; C000595639:erinacine A; C102895:LIMK2 protein, human; C515851:ROCK1 protein, human
keywords: 
  ROS
  ; 
  mTOR
  ; COFL1; PI3K; PROF1; erinacine A; p70S6K