title: [Effect of Nrf2 and TrxR on proliferation of chronic myeloid leukemia cell and its mechanism].
abstract: OBJECTIVE
  To explore the effect of nuclear factor erythroid-2 related factor 2 (Nrf2) and thioredoxin reductase (TrxR) gene on proliferation of chronic myeloid leukemia (CML) line cells and its mechanism.
  
  
  METHODS
  Four interfering sequences of Nrf2 and one negative control sequence were designed and synthesised based on the principle of target sequence of siRNA, then constructed lentivirus vectors, which were transfected into K562 cell lines. The transfection effect was observed by laser scanning confocal microscope (LSCM) and flow cytometer (FCM); The depressing effect of siRNA was analyzed by real-time PCR. The cell proliferation inhibiting rate was measured with CCK-8 assay, the apoptotic rate by Annexin V-PE/PI with FCM and the apoptotic morphology of cells by LSCM.
  
  
  RESULTS
  The transfection efficiency of lentivirus was 65%. One cell line K562-C3 which significantly inhibited Nrf2 mRNA was obtained by real-time PCR, Nrf2 relative quantitation (RQ) expressions were 1.003±0.093 and 0.344±0.032 in the control group and K562-C3 respectively; TrxR expression also decreased with RQ as 1.090±0.549 and 0.395±0.029 respectively. The cellular proliferation inhibition rates of K562-C3 were (4.74±0.39)%, (6.13±1.78)% and (25.36±3.77)%, respectively at 24, 48 and 72 h. The apoptotic rate induced by K562-C3 (29.9%) at 72 hours was obviously higher than in the control group (7.9%). The Annexin V-PE positive K562-C3 cells presented the following apoptotic characteristics, such as karyopyknosis, nuclear fragmentation and apoptotic bodies observed by LSCM.
  
  
  CONCLUSIONS
  Nrf2 specific siRNA could repress its expression at the cellular level and down-regulate the expression of its downstream antioxidant enzyme, such as TrxR, which lead to increased apoptotic rate and decreased cell proliferation.
authors.affiliation: Department of Hematology, 2nd Hospital of Shanxi Medical University, Taiyuan 030001, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Pan
authors.forename: Lian-rong
authors.initials: LR
authors.lastname: Xu
authors.forename: Hong-wei
authors.initials: HW
authors.lastname: Wang
authors.forename: Meng-xia
authors.initials: MX
authors.lastname: Zhu
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.forename: Yan-hong
authors.initials: YH
authors.lastname: Tan
authors.forename: Xiu-hua
authors.initials: XH
authors.lastname: Chen
authors.forename: Fang-gang
authors.initials: FG
authors.lastname: Ren
mesh_terms: D017209:Apoptosis; D049109:Cell Proliferation; D015536:Down-Regulation; D005822:Genetic Vectors; D006801:Humans; D020014:K562 Cells; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000378:metabolism* / Q000473:pathology; D051267:NF-E2-Related Factor 2 / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D013880:Thioredoxin-Disulfide Reductase / Q000378:metabolism*
chemical_list: D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D013880:Thioredoxin-Disulfide Reductase