[Expression of TRAIL gene eukaryotic expression vector modulated by the human telomerase reverse transcriptase gene core promoter in ovarian cancer cell line SKOV3].
OBJECTIVE
To construct the TRAIL gene eukaryotic expression vector modulated by the human telomerase reverse transcription gene core promoter and to evaluate the expression of the TRAIL gene in ovarian cancer cell line SKOV3 in vitro.


METHODS
The amplified TRAIL gene fragment was subsequently cloned into hTERT promoter-pIRES2-EGFP vector and CMVpromoter-pIRES2-EGFP vector. The hTERT promoter-pIRES2-EGFP-TRAIL and CMVpromoter-pIRES2-EGFP-TRAIL eukaryotic expression vectors were obtained, respectively. The plasmids were transfected into human ovarian carcinoma SKOV3 cell line by lipofeclin mediation and the positive clones were screened by G418. The expression of TRAIL gene was examined by RT-PCR, Western blot, immunocytochemistry and flow cytometry.


RESULTS
All the constructed vectors were verified by enzyme digestion. The TRAIL gene of transfected cells was increased significantly (P<0.01).


CONCLUSIONS
An eukaryotic expression vector of the TRAIL gene modulated by the human telomerase reverse transcription gene core promoter has been constructed successfully and its steady expression in human ovarian cancer cell line SKOV3, which will be beneficial to further research into its role in regulating the biological behavior of ovarian cancer cells.
Li
Hong-Mei
HM
Department of Obstetrics and Gynaecology, Affiliated Hospital of Yanan University, Yan'an 716000, China. ylhmqq@yahoo.com.cn
Yu
Yue-Cheng
YC
Song
Tian-Bao
TB
Xin
Xiao-Yan
XY
Zhang
Ming
M
Wei
Xiao-Li
XL
Xu
Jing-Hong
JH
D045744:Cell Line, Tumor; D004262:DNA Restriction Enzymes / Q000378:metabolism; D005057:Eukaryotic Cells / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000473:pathology*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000235:genetics* / Q000378:metabolism; D019098:Telomerase / Q000235:genetics*
D053221:TNF-Related Apoptosis-Inducing Ligand; D019098:Telomerase; D004262:DNA Restriction Enzymes