title: Improved therapeutic effectiveness by combining recombinant p14(ARF) with antisense complementary DNA of EGFR in laryngeal squamous cell carcinoma.
abstract: OBJECTIVE
  The tumor suppressor p14(ARF) and proto-oncogene epidermal growth factor receptor (EGFR) play important roles in the development of laryngeal squamous cell carcinoma (LSCC). This study was aimed to determine whether combining recombinant p14(ARF) with antisense complementary DNA of EGFR could improve the therapeutic effectiveness in LSCC.
  
  
  METHODS
  After human larynx cancer cells (Hep-2) were infected with recombinant adenoviruses (Ad-p14(ARF) and Ad-antisense EGFR) together or alone in vitro, the proliferation and cell cycle distribution of Hep-2 cells were detected by MTT assay and flow cytometer analysis, respectively. Furthermore, the antitumor effects of recombinant adenoviruses together or alone on Hep-2 xenografts were examined in vivo. The levels of p14(ARF) and EGFR expressed in Hep-2 cells and xenografts were determined by western blot assay.
  
  
  RESULTS
  Ad-p14(ARF) combining with Ad-antisense EGFR markedly inhibited the Hep-2 proliferation compared with alone (P=0.001, P=0.002 respectively). Combination of Ad-p14(ARF) and Ad-antisense EGFR led to the proportion of Hep-2 cells in G0/G1 phases increased by up to 86.9%. The down-expression of EGFR protein and overexpression of p14(ARF) protein were observed in vitro and in vivo, and this effect was preserved when Ad-p14(ARF) was combined with Ad-antisense EGFR. Besides, Ad-p14(ARF) plus Ad-antisense EGFR significantly (P<0.05) increased the antitumor activity against Hep-2 tumor xenografts comparing with Ad-p14(ARF) or Ad-antisense EGFR alone.
  
  
  CONCLUSIONS
  Combination Ad-p14(ARF) with Ad-antisense EGFR significantly increased the antitumor responses in LSCC. An effectively potential gene therapy to prevent proliferation of LSCC was provided.
authors.affiliation: West China School of Medicine, West China Hospital, Sichuan University Department of Otorhinolaryngology, Chengdu, People's Republic of China.
authors.forename: Feng
authors.initials: F
authors.lastname: Liu
authors.affiliation: West China School of Medicine, West China Hospital, Sichuan University Department of Otorhinolaryngology, Chengdu, People's Republic of China.
authors.forename: JinTao
authors.initials: J
authors.lastname: Du
authors.affiliation: West China School of Medicine, West China Hospital, Sichuan University Department of Otorhinolaryngology, State Key Laboratory of Biotherapy, Chengdu, People's Republic of China. Electronic address: fxjming@163.com.
authors.forename: Junming
authors.initials: J
authors.lastname: Xian
authors.affiliation: West China School of Medicine, West China Hospital, Sichuan University Department of Otorhinolaryngology, Chengdu, People's Republic of China.
authors.forename: Yafeng
authors.initials: Y
authors.lastname: Liu
authors.affiliation: West China School of Medicine, West China Hospital, Sichuan University Department of Otorhinolaryngology, Chengdu, People's Republic of China.
authors.forename: Shixi
authors.initials: S
authors.lastname: Liu
authors.affiliation: Department of Otorhinolaryngology, the First Affiliated Hospital of Kunming Medical College, Kunming, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Lin
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D002294:Carcinoma, Squamous Cell / Q000473:pathology* / Q000628:therapy; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D016373:DNA, Antisense / Q000235:genetics*; D066246:ErbB Receptors / Q000235:genetics*; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D007822:Laryngeal Neoplasms / Q000473:pathology* / Q000628:therapy; D000090063:Proto-Oncogene Mas; D014162:Transfection; D025803:Tumor Suppressor Protein p14ARF / Q000235:genetics*
chemical_list: D016373:DNA, Antisense; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D025803:Tumor Suppressor Protein p14ARF; D066246:ErbB Receptors