title: [Effect of PML(NLS-) gene mediated by recobinant adenovirus vector on emodin-induced apoptosis of HL-60 cells].
abstract: OBJECTIVE
  To determine the effect and mechanism of action of PML(NLS-) gene on emodin-induced apoptosis of human HL-60 cells.
  
  
  METHODS
  HL-60 cells were infected with recombinant adenovirus Ad-PML (NLS-) and Ad-KZ, respectively. The PML(NLS-) gene was detected by Real-time PCR(RT-PCR) and Western blot. The proliferation level of the HL-60 cells was determined by MTT method. The HL-60 cells were treated with 60 micromol/L emodin for 72 h and then analyzed by flow cytometry for their cell cycle and apoptosis rate. The transcription levels of apoptosis-related BCL-2, BAX and C-MYC genes were determined by RT-PCR. The translation levels of those genes were determined by Western blot.
  
  
  RESULTS
  Compared with normal controls and the HL-60 cells infected with Ad-KZ, the mRNA and protein expression levels of PML(NLS-) gene increased significantly in the HL-60 cells infected with Ad-PML( NLS-). Increased proliferation levels of the Ad-PML (NLS-) infected HL-60 cells were observed in those treated with 60 pmol/L emodin, which showed decreased percentage of cells at Gx phase, increased percentage of cells at S phase, and decreased emodin-induced apoptosis. The levels of mRNA transcription and protein expression of BAX gene decreased, while those of BCL-2 and C-MYC genes increased significantly.
  
  
  CONCLUSIONS
  The over-expression of PML(NLS-) gene might promote the proliferation and arrest the apoptosis of HL-60 cells by up-regulating the expressions of BCL-2 and C-MYC genes and down-regulating the expression of BAX gene.
authors: Central Laboratory of Yong-chuan Hospital Affiliated Chongqing Medical University, Chongqing 402160, China.
authors: Yuan-Mei
authors: YM
authors: Gao
authors: Bei-Zhong
authors: BZ
authors: Liu
authors: Xi
authors: X
authors: Zhang
authors: Xiu-Xiu
authors: XX
authors: Hu
authors: Liang
authors: L
authors: Zhong
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics*; D004642:Emodin / Q000494:pharmacology*; D005260:Female; D005822:Genetic Vectors / Q000235:genetics; D018922:HL-60 Cells; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D000071156:Promyelocytic Leukemia Protein; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: C494826:BAX protein, human; C489427:MYC protein, human; D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; D019253:Proto-Oncogene Proteins c-bcl-2; D016271:Proto-Oncogene Proteins c-myc; D012333:RNA, Messenger; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; D051028:bcl-2-Associated X Protein; C070576:PML protein, human; D004642:Emodin