title: Autophagy regulates Selumetinib (AZD6244) induced-apoptosis in colorectal cancer cells.
abstract: OBJECTIVE
  As Selumetinib is a MEK1/2 inhibitor that has gained interest as an anti-tumor agent, the present study was designed to investigate autophagy involvement on Selumetinib-induced apoptosis in colorectal cancer (CRC) cells.
  
  
  METHODS
  CRC cells death and cycle studies were assessed by AnnexinV-FITC and PI staining, respectively. Autophagy flux was analysed by Western Blot (LC3II and p62 protein levels) and retroviral infection of SW480 cells for siBecn1 RNA interference experiments. Confocal microscopy was used to determine mCherry-EGFP-LC3 distribution.
  
  
  RESULTS
  The Selumetinib effects were concentration-dependent in SW480 cell line. Whereas 1 μM exerted an arrest in the cell cycle (G1 phase), higher concentrations (10 μM) induced cell death, which was accompanied by autophagy blockage in its last stages. Autophagy induction by Rapamycin (RAPA) increased cell survival, whereas pharmacology autophagy inhibition by Bafilomycin A1 (BAF), Chloroquine (CQ) or 3-Methyladenine (3-MA) increased Selumetinib-induced CRC cells death.
  
  
  CONCLUSIONS
  Altogether, these results suggest that autophagy plays a fundamental role in CRC cells response to Selumetinib. In addition, the combination of Selumetinib with autophagy inhibitors may be a useful therapeutic strategy to enhance its activity against colorectal tumours.
authors.affiliation: Instituto de Biología Molecular y Celular, Universidad Miguel Hernández, 03202 Elche, Alicante, Spain; Departamento de Farmacologia, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, SP, Brazil.
authors.forename: Silvina
authors.initials: S
authors.lastname: Grasso
authors.affiliation: Departamento de Farmacologia, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, SP, Brazil.
authors.forename: Gustavo J S
authors.initials: GJS
authors.lastname: Pereira
authors.affiliation: Departamento de Farmacologia, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, SP, Brazil.
authors.forename: Caroline
authors.initials: C
authors.lastname: Palmeira-Dos-Santos
authors.affiliation: Departamento de Farmacologia, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, SP, Brazil.
authors.forename: Andrana K
authors.initials: AK
authors.lastname: Calgarotto
authors.affiliation: Instituto de Biología Molecular y Celular, Universidad Miguel Hernández, 03202 Elche, Alicante, Spain; Unidad AECC de Investigación Traslacional en Cáncer, Hospital Universitario Virgen de la Arrixaca, Instituto Murciano de Investigación Biosanitaria, 30120 Murcia, Spain.
authors.forename: Isabel
authors.initials: I
authors.lastname: Martínez-Lacaci
authors.affiliation: Instituto de Biología Molecular y Celular, Universidad Miguel Hernández, 03202 Elche, Alicante, Spain.
authors.forename: Jose Antonio
authors.initials: JA
authors.lastname: Ferragut
authors.affiliation: Departamento de Farmacologia, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, SP, Brazil. Electronic address: soraya.smaili23@gmail.com.
authors.forename: Soraya S
authors.initials: SS
authors.lastname: Smaili
authors.affiliation: Departamento de Farmacologia, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, SP, Brazil. Electronic address: claudia.bincoletto@unifesp.br.
authors.forename: Claudia
authors.initials: C
authors.lastname: Bincoletto
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001343:Autophagy / Q000187:drug effects*; D001562:Benzimidazoles / Q000737:chemistry / Q000494:pharmacology*; D002453:Cell Cycle / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D015179:Colorectal Neoplasms / Q000473:pathology*; D019073:HT29 Cells; D006801:Humans
chemical_list: C517975:AZD 6244; D000970:Antineoplastic Agents; D001562:Benzimidazoles
keywords: Apoptosis; Autophagy; CRC cells (SW480 and HT29); Selumetinib (AZD6244)