title: [Construction of TRAIL gene eukaryotic expression vector modulated by hTERT gene core promoter and its effect on apoptosis of ovarian cancer cells].
abstract: OBJECTIVE
  To construct the TNF-related apoptosis inducing ligand(TRAIL) gene eukaryotic expression vector modulated by human telomerase reserse transcriptase (hTERT) gene core promoter and to study its effect on apoptosis of ovarian cancer cells.
  
  
  METHODS
  Genomic RNA was extracted from human placenta tissues and the fragment of TRAIL was obtained by RT-PCR. The amplified gene fragment was subsequently cloned into hTERTpromoter-pIRES2-EGFP vector and CMV promoter-pIRES2-EGFP vector after sequencing. The hTERTpromoter-pIRES2-EGFP-TRAIL and CMV promoter-pIRES2-EGFP-TRAIL eukaryotic expression vectors were constructed respectively. The recombinant plasmids were transfected into ovarian carcinoma cell line, SKOV3, and the levels of mRNA were determined by RT-PCR. The cell cycle and apoptosis rate of SKOV3 cells were determined by FCM.
  
  
  RESULTS
  The constructed two recombinant vectors were verified by restriction enzyme digestion analysis and DNA sequencing. After being transfected with two recombinant vectors, the growth of SKOV3 cells was strongly inhibited and apoptotic features appeared.
  
  
  CONCLUSIONS
  The recombinant eukaryotic expression vector has been constructed successfully. TRAIL gene driven by hTERTpromoter can be obviously expressed in ovarian carcinoma SKOV3 cells, suggesting that the specific expression vector modulated by hTERT core gene promoter may be a novel and promising approach to the tumor treatment.
authors.affiliation: Environment and Genes Related to Diseases Key Laboratory of Education Ministry, School of Medicine, Xi'an Jiaotong University, Xi'an 710061, China. ylhmqq@yahoo.com.cn
authors.forename: Hong-mei
authors.initials: HM
authors.lastname: Li
authors.forename: Tian-bao
authors.initials: TB
authors.lastname: Song
authors.forename: Yue-cheng
authors.initials: YC
authors.lastname: Yu
authors.forename: Yun-xin
authors.initials: YX
authors.lastname: Cao
authors.forename: Hong-yin
authors.initials: HY
authors.lastname: Wang
authors.forename: Hui
authors.initials: H
authors.lastname: Song
authors.forename: Ming
authors.initials: M
authors.lastname: Zhang
mesh_terms: D017209:Apoptosis / Q000235:genetics*; D005260:Female; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D010051:Ovarian Neoplasms / Q000473:pathology* / Q000628:therapy; D010957:Plasmids; D011401:Promoter Regions, Genetic; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000235:genetics* / Q000378:metabolism; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D053221:TNF-Related Apoptosis-Inducing Ligand; C509186:TERT protein, human; D019098:Telomerase