title: Effect of capilliposide for induction apoptosis in human nasopharyngeal cancer CNE-2 cells through up-regulating PUMA expression.
abstract: OBJECTIVE
  To observe the apoptosis of capilliposide against human nasopharyngeal cancer CNE-2 cells and to study its primary mechanisms.
  
  
  METHODS
  Vectors pSilencer-PUMA-small interfering RNA (siRNA) were constructed to transcribe functional siRNA specially targeting PUMA. The interfering plasmids were used to transfect CNE-2 cells with lipofectamine 2000 transfection reagent. PUMA messenger RNA (mRNA) expression levels were analyzed by polymerase chain reaction. The proliferation of CNE-2 cells was detected using MTT colorimetry. Annexin V/propidium iodide double staining was applied to detect the apoptosis rate of CNE-2 cells. The protein levels of p53, PUMA, and Bax were detected using Western blot analysis.
  
  
  RESULTS
  Recombinant siRNA expression vector targeting PUMA was constructed. MTT assays showed capilliposide inhibited the proliferation of CNE-2 cells in a concentration-dependent manner. The inhibition was strengthened along with increased concentrations. Apoptosis detected by flow cytometry in control group, drug group, siRNA group, and drug combined siRNA group was 9.3 ± 2.3%, 31.4 ± 5.6%, 12.3 ± 4.1%, and 13.2 ± 3.7%, respectively. After pretreated by capilliposide, PUMA protein was upregulated, and BAX was distributed to mitochondria in CNE-2 cells using Western blot analysis, but this effect can be interrupted by PUMA-siRNA.
  
  
  CONCLUSIONS
  Capilliposide could induce the apoptosis of CNE-2 cells, which might be related with the increasing in PUMA-Bax pathway.
authors.forename: Yonghong
authors.initials: Y
authors.lastname: Hua
authors.affiliation: Department of Radiation Oncology, Zhejiang Cancer Hospital, Hangzhou 310022, China.
authors.forename: Qiaoying
authors.initials: Q
authors.lastname: Hu
authors.forename: Yongfeng
authors.initials: Y
authors.lastname: Piao
authors.forename: Qiu
authors.initials: Q
authors.lastname: Tang
authors.forename: Jiangguo
authors.initials: J
authors.lastname: Feng
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004354:Drug Screening Assays, Antitumor; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D009303:Nasopharyngeal Neoplasms / Q000188:drug therapy*; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D012503:Saponins / Q000494:pharmacology*; D014315:Triterpenes / Q000494:pharmacology*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D051017:Apoptosis Regulatory Proteins; C494826:BAX protein, human; C438378:BBC3 protein, human; D011518:Proto-Oncogene Proteins; D012503:Saponins; C495901:TP53 protein, human; D014315:Triterpenes; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein