title: Effect of silencing key proteins in telomerase mechanism and alternative lengthening of telomeres mechanism in laryngeal cancer cells.
abstract: OBJECTIVE
  To explore the influences of telomerase and alternative lengthening of telomeres mechanism on telomere length and laryngeal squamous cell carcinoma in vitro and in vivo.
  
  
  METHODS
  Short hairpin RNA expression vectors targeting the messenger TERT, TRF2, RAD51 and NBS1 were constructed. The mRNA and protein expression of targeted genes in human laryngeal squamous carcinoma cell line HEp-2 was evaluated by reverse transcription polymerase chain reaction and Western blotting separately. The length of telomere was analyzed by fluorescent in-situ hybridization. Cell viability was examined by cell counting Kit-8. Effects on tumor growth were also investigated in vivo.
  
  
  RESULTS
  The transfection of multiple short hairpin RNAs expression plasmid significantly inhibited the mRNA and protein expression of related genes. Silence of alternative lengthening of telomeres mechanism and telomerase mechanism related genes resulted in the shortening of telomere length in HEp-2 cell. However, silence of alternative lengthening of telomeres mechanism related genes could shorten the telomere length but had no significant difference. Both simultaneously and separately blocking telomerase mechanism and alternative lengthening of telomeres mechanism resulted in reduction of tumor cell viability. Silence of alternative lengthening of telomeres mechanism and telomerase mechanism related genes inhibited the tumor growth in vivo.
  
  
  CONCLUSIONS
  The inhibition of telomere related gene may be a promising strategy for the treatment of laryngeal squamous cell carcinoma.
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Ultrasonography, Zhongnan Hospital of Wuhan University, Wuhan, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan, China.
authors.forename: Shi-Ming
authors.initials: SM
authors.lastname: Chen
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Chen
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan, China.
authors.forename: Bo-Kui
authors.initials: BK
authors.lastname: Xiao
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan, China. Electronic address: taozezhang@hotmail.com.
authors.forename: Ze-Zhang
authors.initials: ZZ
authors.lastname: Tao
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000473:pathology*; D018929:Cell Culture Techniques; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival; D006801:Humans; D007822:Laryngeal Neoplasms / Q000378:metabolism / Q000473:pathology*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051135:Rad51 Recombinase / Q000235:genetics / Q000378:metabolism; D019098:Telomerase / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D016615:Telomere; D059505:Telomere Homeostasis / Q000502:physiology*; D035341:Telomeric Repeat Binding Protein 2 / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D018797:Cell Cycle Proteins; C112156:NBN protein, human; D009687:Nuclear Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C504753:TERF2 protein, human; D035341:Telomeric Repeat Binding Protein 2; D051135:Rad51 Recombinase; C509186:TERT protein, human; D019098:Telomerase