title: [Inhibitory effect of Rnai on AML1 -ETO fusion gene expression in leukemia cells].
abstract: OBJECTIVE
  By inhibiting AML1 -ETO fusion gene expression in Kasumi-1 cells with RNAi, to investigate the changes in cell proliferation and cell cycle.
  
  
  METHODS
  The small interference RNAs (siRNAs) specifically targeting the AML1 -ETO fusion gene were synthesized in vitro and transfected into Kasumi-1 cells by electroporation, the non-specific siRNAs transfected cells were taken as control. EGFP plasmid was transfected into Kasumi-1 cell and the transfection efficiency was detected by FCM. Inhibitory effect of siRNAs were detected by real-time RT-PCR and Western blots. Cell proliferation was measured by CCK-8 assay. DNA content was detected by PI assay.
  
  
  RESULTS
  The transfection efficiency was 44.5%. The AML1 -ETO specific siRNAs inhibited AML1 -ETO expression at both mRNA and protein levels. The cell proliferation rate in siRNAs treated group was lower than that in control group 72 h after transfection [(47.90 +/- 0.02)% vs (66.90 +/- 0.08)% , P < 0.05]. The cell cycle was blocked at G1 phase 72 h after siRNAs treatment, the cell proportion in G1 phase being 38.3% and 31.6% in control group, while in G2/M phase being 1.8% and 2.4% respectively.
  
  
  CONCLUSIONS
  The synthesized siRNAs can inhibit AML1 -ETO fusion gene expression. AML1 -ETO specific siRNA induced the decline of AML1 -ETO fusion protein in Kasumi-1 cell, and then caused the cell cycle blocked in G1 stage and eventually inhibited the cell proliferation.
authors.affiliation: Department of Hematology, Shanghai First People's Hospital, Shanghai 200080, China.
authors.forename: Ju
authors.initials: J
authors.lastname: Wei
authors.forename: Su
authors.initials: S
authors.lastname: Li
authors.forename: Chun
authors.initials: C
authors.lastname: Wang
authors.forename: You-Wen
authors.initials: YW
authors.lastname: Qin
authors.forename: Xiao-Xia
authors.initials: XX
authors.lastname: Ma
authors.forename: Kuang-Cheng
authors.initials: KC
authors.lastname: Xie
authors.forename: Shi-Ke
authors.initials: SK
authors.lastname: Yan
authors.forename: Yan-Rong
authors.initials: YR
authors.lastname: Gao
authors.forename: Qi
authors.initials: Q
authors.lastname: Cai
mesh_terms: D002453:Cell Cycle / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D050676:Core Binding Factor Alpha 2 Subunit / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007938:Leukemia / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D015514:Oncogene Proteins, Fusion / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D000075142:RUNX1 Translocation Partner 1 Protein; D014162:Transfection
chemical_list: C107844:AML1-ETO fusion protein, human; D050676:Core Binding Factor Alpha 2 Subunit; D015514:Oncogene Proteins, Fusion; D000075142:RUNX1 Translocation Partner 1 Protein