[Construction of TRAIL gene eukaryotic expression vector modulated by hTERT gene core promoter and its effect on apoptosis of ovarian cancer cells].
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.
Li
Hong-mei
HM
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
Song
Tian-bao
TB
Yu
Yue-cheng
YC
Cao
Yun-xin
YX
Wang
Hong-yin
HY
Song
Hui
H
Zhang
Ming
M
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
D053221:TNF-Related Apoptosis-Inducing Ligand; C509186:TERT protein, human; D019098:Telomerase