[Inhibitory effect of antisense human telomerase reverse transcriptase(hTERT) on telomerase activity of human pulmonary giant cell carcinoma cell line(PLA-801D)].
OBJECTIVE
Telomerase has been thought to play an important role in the carcinogenesis in recent years. Human telomerase reverse transcriptase (hTERT) is a limiting component for telomerase activity. This study was designed to explore the effect of transfection of the full-length cDNA of antisense hTERT on the malignant phenotype of human pulmonary giant cell carcinoma cell line (PLA-801D) and its potential role in the gene therapy for cancers.


METHODS
An antisense hTERT cDNA eukaryotic expression vector pcDNA3.1(-)-hTERT including the full length of hTERT cDNA sequence was constructed using recombinant DNA technique and transfected into human pulmonary giant cell carcinoma cells (PLA-801D) with liposome. The effect of antisense hTERT on the cellular proliferation capacity of PLA-801D cells was analyzed by the growth curve. The expression of hTERT mRNA was examined by reverse transcription polymerase chain reaction (RT-PCR). The telomerase activity was determined by telomeric-repeat amplification protocol enzyme-linked immunoassay (TRAP-ELISA).


RESULTS
Antisense pcDNA3.1 (-)-hTERT eukaryotic expression have been constructed and was successfully transfected into the PLA-801D cells. The growth speed of PLA-801D transfected with antisense hTERT was significantly inhibited compared with the control cells, and the hTERT mRNA expression was inhibited, the relatively expression was only 15.7% of control cells, and telomerase activity was down-regulated about 82.4%.


CONCLUSIONS
Full-length antisense hTERT cDNA can suppress hTERT mRNA expression and telomerase activity, and restrict the growth speed of tumor cells.
Ma
Xue-Bin
XB
Molecular Biology Laboratory, General Hospital of PLA, Beijing, 100853, PR China.
Han
Wei-Dong
WD
Hao
Hao-Jie
HJ
Li
Qi
Q
Xu
Zhou-Min
ZM
Lu
Xue-Chun
XC
Zhao
Ya-Li
YL
D018286:Carcinoma, Giant Cell / Q000201:enzymology* / Q000628:therapy; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins; D004797:Enzyme-Linked Immunosorbent Assay; D015316:Genetic Therapy; D006801:Humans; D008175:Lung Neoplasms / Q000201:enzymology* / Q000628:therapy; D016372:RNA, Antisense / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019098:Telomerase / Q000037:antagonists & inhibitors* / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection
D004268:DNA-Binding Proteins; D016372:RNA, Antisense; D012333:RNA, Messenger; D019098:Telomerase