Effect of PML(NLS-) Gene Mediated by Recobinant Adenovirus Vector on Emodin-induced Apoptosis of HL-60 Cells

GAO Yuan-mei, LIU Bei-zhong, ZHANG Xi, HU Xiu-xiu, ZHONG Liang

Abstract

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 μmol/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 μmol/L emodin, which showed decreased percentage of cells at G1 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. Conclusion 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.

 

Keywords: PML(NLS-) gene, Leukemia, HL-60 cells, Apoptosis

 

Full Text:

PDF


References


Lane АЛ, Ley TJ. Neutrophil elastase is important for PML- retinoic acid receptor a activities in early myeloid cells. Mol Cell Biol ,2005; 25(1); 23-33.

Leung D. Adenoviridae; gene therapy and the molecular biology of the adenovirus. The Meducator,2012; 1( 1): 15-16.

Tallman MS. Treatment of relapsed or refractory acute promyelocytic leukemia. Best Pract Res Clin Haematol.2007;20 (1): 57-65.

Wang L, Zhou GB, Liu P, el al. Dissection of mechanisms of Chinese medicinal formula Realgar-Indigo naturalis as an effective treatment for promyelocytic leukemia. PNAS.2008; 105 ( 12):4 826-4831.

Zhou GB, Zhang J, Wang ZY, et al. Treatment of acute promyelocytic leukaemia with all-trans retinoic acid and arsenic trioxide: a paradigm of synergistic molecular targeting therapy. Phil Trans R Soc B,2007;362( 1482) :959-971.

Krieghoff-Henning E, Hofmann TG. Role of nuclear bodies in apoptosis signalling. Biochim Biophys Acta, 2008; 1783 ( 11 ): 2185-2194.

Bernardi R, Papa A, Pandolfi PP. Regulation of apoptosis by PML and the PML-NBs. Oncogene,2008;27(48) :6299-6312.

Shima Y, Shima T, Chiba T, et al. PML activates transcription by protecting HIPK2 and p300 from SCFFbx3- mediated degradation. Mol Cell Biol,2008;28(23):7126-7138.


Refbacks

  • There are currently no refbacks.