[Inhibitory effect of antisense human telomerase reverse transcriptase(hTERT) on telomerase activity of human pulmonary giant cell carcinoma cell line(PLA-801D)].

Abstract

BACKGROUND & 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%.

CONCLUSION:

Full-length antisense hTERT cDNA can suppress hTERT mRNA expression and telomerase activity, and restrict the growth speed of tumor cells.

• Authors: Xue-Bin Ma, Wei-Dong Han, Hao-Jie Hao, Qi Li, Zhou-Min Xu, Xue-Chun Lu, Ya-Li Zhao
• Journal: Ai zheng = Aizheng = Chinese journal of cancer (Ai Zheng) Vol. 22 Issue 9 Pg. 932-7 (Sep 2003) China
• PMID: 12969524 (Publication Type: English Abstract, Journal Article)

Chemical References

• DNA-Binding Proteins
• RNA, Antisense
• RNA, Messenger
• Telomerase

Topics

• Carcinoma, Giant Cell (enzymology, therapy)
• Cell Line, Tumor
• DNA-Binding Proteins
• Enzyme-Linked Immunosorbent Assay
• Genetic Therapy
• Humans
• Lung Neoplasms (enzymology, therapy)
• RNA, Antisense (pharmacology)
• RNA, Messenger (analysis)
• Reverse Transcriptase Polymerase Chain Reaction
• Telomerase (antagonists & inhibitors, genetics, metabolism)
• Transfection