title: RNA interference with EAG1 enhances interferon gamma injury to glioma cells in vitro.
abstract: OBJECTIVE
  The aim of this study was to silence Ether à go-go 1 (EAG1) in glioma cells by RNAi in order to further analyze whether silencing this channel would improve injury caused by interferon gamma (IFN-γ).
  
  
  METHODS
  EAG1 silencing by the siRNAs EAG1hum_287 and EAG1hum_1727 (sequence targets 5'-GGCCTATTGTGTACAGCAATT-3' and 5'-GGGACTTCCTGAAGCTCTATT-3', respectively) was determined by reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR). Cell viability was measured by the 3-(4,5)-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) assay. U-138MG glioma cells were injured by IFN-γ (25 ng/ml, 24 h) with or without the RNAi for EAG1 by a non-viral vector (pKV10.1-3, 0.2 μg).
  
  
  RESULTS
  EAG1hum_287 and EAG1hum_1727 caused 0.46- and 0.52-fold decrease in EAG1 mRNA content, respectively. RNAi for EAG1 by pKv10.1-3 strengthened the reduction in cell viability caused by IFN-γ (11.4% versus 40.4%, p<0.05).
  
  
  CONCLUSIONS
  The present study reinforces the notion that EAG1 has a role in glioma biology, suggesting that this channel is a relevant player preserving the cell viability during IFN-γ injury.
authors.affiliation: Technology for Gene Therapy Laboratory, University of Brasilia - UnB/FAV, Darcy Ribeiro campus, ICC - ASS 128, Brasília, DF, Brazil. ricardo.titze@hotmail.com
authors.forename: Ludmylla Costa
authors.initials: LC
authors.lastname: Cunha
authors.forename: Elaine
authors.initials: E
authors.lastname: Del Bel
authors.forename: Luis
authors.initials: L
authors.lastname: Pardo
authors.forename: Walter
authors.initials: W
authors.lastname: Stühmer
authors.forename: Ricardo
authors.initials: R
authors.lastname: Titze-DE-Almeida
mesh_terms: D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D051638:Ether-A-Go-Go Potassium Channels / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D005910:Glioma / Q000473:pathology*; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics
chemical_list: D051638:Ether-A-Go-Go Potassium Channels; C496884:KCNH1 protein, human; D034741:RNA, Small Interfering; D007371:Interferon-gamma