title: [Construction and selection of siRNA expression cassettes targeting human telomerase reverse transcriptase gene in vitro].
abstract: OBJECTIVE
  To determine whether the human telomerase reverse transcriptase (hTERT) gene silencing could be effectively induced by PCR-derived siRNA expression cassettes (SEC) transfected by the fifth generation polyamidoamine dendrimer (G5 PAMAM-D) in Tca8113 cells.
  
  
  METHODS
  Four SEC were rationally designed and constructed based on a two-step PCR reaction. The SEC were then transferred into Tca8113 cells using G5 PAMAM-D, and hTERT expression was investigated by real-time fluorescence-quantitative reverse transcriptase-PCR and western blot analysis.
  
  
  RESULTS
  The RNA interference effects of the SEC targeted for varying hTERT mRNA positions showed a significant disparity. Among them, SEC-A revealed the most potent inhibitory effects (above 95% of reduction), followed by SEC-D and SEC-C, and SEC-B had no effect on hTERT expression (P > 0.05). That the endogenous hTERT gene silencing induced by G5 PAMAM dendrimer-mediated SEC-A was highly sequence-specific, and multiple transfection as well as properties of the vectors were routinely attributable to the specific suppression.
  
  
  CONCLUSIONS
  Specific inhibition of endogenous hTERT expression by use of a PCR-based short hairpin siRNA technique and dendrimer transfer system may serve as a novel strategy for treatment of tongue cancers expressing hTERT in vitro.
authors.affiliation: Department of Oral and Maxillofacial Surgery, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou 510055, China.
authors.forename: Xi-qiang
authors.initials: XQ
authors.lastname: Liu
authors.forename: Hong-zhang
authors.initials: HZ
authors.lastname: Huang
authors.forename: Chao-bin
authors.initials: CB
authors.lastname: Pan
authors.forename: Fang-jin
authors.initials: FJ
authors.lastname: Wang
authors.forename: Bin
authors.initials: B
authors.lastname: Zhang
authors.forename: Wei-wen
authors.initials: WW
authors.lastname: Liang
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000201:enzymology* / Q000235:genetics; D045744:Cell Line, Tumor; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D034741:RNA, Small Interfering / Q000235:genetics*; D019098:Telomerase / Q000235:genetics*; D014062:Tongue Neoplasms / Q000201:enzymology* / Q000235:genetics; D014162:Transfection
chemical_list: D034741:RNA, Small Interfering; C509186:TERT protein, human; D019098:Telomerase