Adenovirus-mediated HSV-TK gene therapy using the human telomerase promoter induced apoptosis of small cell lung 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, but 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 tested the possibility of the utilization of the hTERT promoter in targeted cancer gene therapy. We cloned the hTERT promoter in the place 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 a normal and a small cell lung cancer cell line. Through these experiments, we identified the selective tumor specific cell death by Ad-hT-TK. Furthermore, cell death detection ELISA and FACS analysis suggest 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 SCLC gene therapy.
Song
Joon-Seok
JS
Institute of Biotechnology, Korea University, Seoul 136-701, Korea. antibody@korea.ac.kr
Kim
Hyun-Pyo
HP
D000256:Adenoviridae / Q000235:genetics*; D017209:Apoptosis; D018288:Carcinoma, Small Cell / Q000235:genetics / Q000473:pathology* / Q000628:therapy*; D002384:Catalysis; D002460:Cell Line; D045744:Cell Line, Tumor; D002469:Cell Separation; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000378:metabolism; D005434:Flow Cytometry; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008175:Lung Neoplasms / Q000235:genetics / Q000473:pathology* / Q000628:therapy*; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011505:Protein-Tyrosine Kinases / Q000235:genetics*; D018139:Simplexvirus / Q000235:genetics*; D019098:Telomerase / Q000235:genetics*; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D008156:Luciferases; D011505:Protein-Tyrosine Kinases; D019098:Telomerase