title: [Phosphorylation status of ASPP2 modulates p53 apoptotic function in oxaliplatin-induced apoptosis of colorectal cancer HCT116 cells].
abstract: OBJECTIVE
  To investigate the role of apoptosis stimulating protein 2 of p53 (ASPP2) phosphorylation status in the regulation of ASPP2-p53 apoptotic pathway activity.
  
  
  METHODS
  Cells were individually transfected with green fluorescent protein (GFP)-encoding vector, constitutively non-phosphorylatable ASPP2 mutant-ASPP2 (Am)-encoding vector, and wild type ASPP2 (Aw)-encoding vector) plasmids, respectively, to make them overexpressing phosphorylated and non-phosphorylated ASPP2 proteins, respectively. Cell apoptosis was induced by oxaliplatin. The apoptosis rate of cells was determined by flow cytometry after staining with FITC-conjugated annexin V and PI. ASPP2 protein level and its phosphorylation status were observed by Western blot. The interaction between ASPP2 and p53 was observed by immunoprecipitation assay.
  
  
  RESULTS
  Oxaliplatin induced cell apoptosis and caused phosphorylation of ASPP2 at ser92/ser361 in the HCT116 cells. The apoptosis rate of Aw and Am plasmids-transfected cells were (3.8 ± 1.0)% and (3.9 ± 1.2)% respectively, statistically with a non-significant difference (P > 0.05) in comparison with that of the GFP plasmid-transfected cells [(4.0 ± 0.8)%]. After oxaliplatin treatment, the apoptosis rate of Aw plasmid-transfected cells was (46.7 ± 3.9)%, significantly higher than that of the Am and GFP plasmid-transfected cells [(40.1 ± 10.2)% and (37.1 ± 6.9)%, respectively, P < 0.05], however, there was no statistically significant difference (P > 0.05) between Am and GFP plasmid-transfected cells. These results indicate that phosphorylated ASPP2 promoted the oxaliplatin-induced apoptosis of HCT116 cells through a p53-dependent pathway. Phosphorylation status of ASPP2 influenced its binding activity to p53.
  
  
  CONCLUSIONS
  Phosphorylation status of ASPP2 modulates p53 apoptotic function in oxaliplatin-induced apoptosis of colorectal cancer HCT116 cells.
authors.affiliation: Department of General Surgery, Cancer Hospital of Shandong Province, Jinan 250117, China.
authors.forename: Qingsheng
authors.initials: Q
authors.lastname: Hou
authors.forename: Hongwei
authors.initials: H
authors.lastname: Zhao
authors.forename: Weipeng
authors.initials: W
authors.lastname: Gong
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Zhu
authors.forename: Yue
authors.initials: Y
authors.lastname: Han
authors.forename: Dexi
authors.initials: D
authors.lastname: Chen
authors.affiliation: Email: hliangbb@163.com.
authors.forename: Hongliang
authors.initials: H
authors.lastname: Guo
mesh_terms: D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D015179:Colorectal Neoplasms / Q000378:metabolism*; D045325:HCT116 Cells; D006801:Humans; D009944:Organoplatinum Compounds; D000077150:Oxaliplatin; D010766:Phosphorylation; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*
chemical_list: D051017:Apoptosis Regulatory Proteins; D009944:Organoplatinum Compounds; C090468:TP53BP2 protein, human; D016159:Tumor Suppressor Protein p53; D000077150:Oxaliplatin