title: [Effect of ginseng polysaccharide-induced wnt/beta-catenin signal transduction pathway on apoptosis of human nasopharyngeal cancer cells CNE-2].
abstract: OBJECTIVE
  To observe the effect of ginseng polysaccharide (GPS) on the proliferation and apoptosis of human nasopharyngeal cancer cells CNE-2, and discuss the possible mechanism.
  
  
  METHODS
  The effect of GPS on the growth of CNE-2 cells was observed by CCK8 assay. CNE-2 cells in the logarithmic phase were collected and processed respectively with different concentrations (0, 0. 1, 0. 2, 0. 3. 0. 4 g L-1) of GPS for 48 h. The flow cytometry was used to detect its induction effect on CNE-2 cell apoptosis. Hoechst-33258 cell staining and electron microscope were used to observe the morphological changes of cells. The beta-catenin mRNA expression was detected by Real-time PCR. The protein localizations and expressions of beta-catenin and TCF4 were tested by the immunofluorescence staining. The expressions of beta-catenin, Bcl-2 and Bax proteins were detected by Western blot.
  
  
  RESULTS
  CCK8 assay results showed that GPS could remarkably inhibit the proliferation of CNE-2 cells, with dose-time dependence. IC50 of cells induced with GPS for 48 h was 0. 39 g L-1. After being processed with GPS with concentrations of 0.1, 0. 2, 0. 3, 0. 4 g L-1 for 48 h, the cell apoptosis rates of human nasopharyngeal cancer cells CNE-2 were (5. 69 +/- 0. 29)% , (10. 3 +/- 0. 63)% , (15. 4 +/- 0. 74 ) % and (35. 7 +/- 1. 86)% , respectively. Significant difference was observed compared with the control group (2. 08 +/- 0. 11) % (P <0. 05). The results of Hoechst-33258 staining showed the characteristics of cell apoptosis. Under the electron microscope, apoptosis bodies could be observed among CNE-2 cells induced with GPS with the concentration of 0. 4 g L -1 for 48 h. The results of Real-time PCR showed a significant reduction in beta-catenin mRNA expression. The results of laser confocal microscopy revealed notable decrease of beta-catenin and TCF4 expression in nucleus and transfer from nucleus to cell membranes in beta-catenin expression areas after being processed with GPS for 48 h. Western blot showed significant decrease in the expressions of beta-catenin and anti-apoptosis protein Bcl-2, with an increasing expression in apoptosis-promoting protein Bax (P <0. 05).
  
  
  CONCLUSIONS
  GPS could significantly inhibit the proliferation of CNE-2 cells and promote thier apoptosis. The obstruction of Wnt/beta-catenin signaling pathway may be an important mechanism for GPS to induce the apoptosis of human nasopharyngeal cancer cells CNE-2.
authors.affiliation: Department of Otorhinolaryngology-Head and Neck Surgery, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China. fanwhich@163.com
authors.forename: Jia-Ming
authors.initials: JM
authors.lastname: Fan
authors.affiliation: Laboratory of Stem Cell and Tissue Engineering, Teaching and Research Room of Histology and Embryology, S Chongqing Medical University, Chongqing 400016, China.
authors.forename: Ze-Hong
authors.initials: ZH
authors.lastname: Liu
authors.affiliation: Laboratory of Stem Cell and Tissue Engineering, Teaching and Research Room of Histology and Embryology, S Chongqing Medical University, Chongqing 400016, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: Laboratory of Stem Cell and Tissue Engineering, Teaching and Research Room of Histology and Embryology, S Chongqing Medical University, Chongqing 400016, China.
authors.forename: Ya-Ping
authors.initials: YP
authors.lastname: Wang
authors.affiliation: Department of Otorhinolaryngology-Head and Neck Surgery, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: Lv-Yuan
authors.initials: LY
authors.lastname: Yang
authors.affiliation: Department of Otorhinolaryngology-Head and Neck Surgery, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: Jiang-Ju
authors.initials: JJ
authors.lastname: Huang
mesh_terms: D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D002277:Carcinoma; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D006801:Humans; D000077274:Nasopharyngeal Carcinoma; D009303:Nasopharyngeal Neoplasms / Q000378:metabolism*; D005894:Panax / Q000737:chemistry*; D011134:Polysaccharides / Q000494:pharmacology*; D060888:Real-Time Polymerase Chain Reaction; D060449:Wnt Signaling Pathway / Q000187:drug effects; D051176:beta Catenin / Q000235:genetics
chemical_list: D011134:Polysaccharides; D051176:beta Catenin