Adenovirus-mediated suicide gene therapy using the human telomerase catalytic subunit (hTERT) gene promoter induced apoptosis of ovarian cancer cell line.
Telomerase is a ribonucleoprotein complex the function of which is to add telomeric repeats (TTAGGG)(n) to chromosomal ends, and it is known to play an important role in cellular immortalization. Telomerase is highly active in most tumor cells, yet not in normal cells. As such, it may have possible applications in cancer gene therapy. Telomerase consists of two essential components, telomerase RNA template (hTR) and catalytic subunit (hTERT). hTERT is expressed only in cells and tissues positive for telomerase activity, i.e., tumor and fetal cells. We here tested the possibility of the utilization of the hTERT promoter in targeted cancer gene therapy. We cloned the hTERT promoter in the replace of the CMV promoter and sub-cloned HSV-TK gene to be controlled by hTERT gene promoter in adenovirus shuttle plasmid. Then we constructed recombinant adenovirus Ad-hT-TK, and infected them into normal and human gynecological cancer cell lines. Through these experiments, we identified the selective tumor specific cell death by Ad-hT-TK. Furthermore, FACS analysis and TUNEL assay suggests that the reduced viability is mediated through the induction of apoptosis, indicating that this approach may be a useful method for suppressing cancer growth in targeted cancer gene therapy. These results show that Ad-hT-TK could be used for gynecological cancer gene therapy.
Song
Joon-Seok
JS
Institute of Biotechnology, Korea University, Seoul, Korea.
Kim
Hyun-Pyo
HP
Yoon
Won-Suck
WS
Lee
Kyu-Wan
KW
Kim
Mee-Hye
MH
Kim
Kyung-Tai
KT
Kim
Hy-Sook
HS
Kim
Young Tae
YT
D000256:Adenoviridae / Q000235:genetics*; D017209:Apoptosis; D002460:Cell Line; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins; D005260:Female; D005347:Fibroblasts / Q000201:enzymology; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D010051:Ovarian Neoplasms / Q000628:therapy*; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D021122:Protein Subunits / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019098:Telomerase / Q000235:genetics*
D004268:DNA-Binding Proteins; D021122:Protein Subunits; D008156:Luciferases; D019098:Telomerase