Antisense human telomerase reverse transcriptase repress the expression of p53 of human pulmonary giant cell carcinoma cell line

Objective:In this study, the authors investigated the effects of the full length of antisense human telomerase reverse transcriptase （hTERT） gene on the expression of p53 and the distrubution of cell cycles of human pulmonary giant cell carcinoma cell line （PLA-801D）. Methods:An antisense hTERT gene eukaryotic expression vector pcDNA 3.1（-）-hTERT including the full-length region of hTERT CDS was constructed by recombinant DNA technique and transfected into the human pulmonary giant cell carcinoma cell line （PLA-801D）. Then the expression of p53 mRNA was observed with RT-PCR, distrubution of cell cycles was determined by flow cytometry. The expression of mutated p53 protein was tested by immunohischemitry. Results:Full length （3.4kb） of hTERT fragment was inserted into vector pcDNA 3.1（-） in reverse orientation have been constructed and was transfected successfully into the PLA-801D cells, the distribution of cell cylce of antisense hTERT transfected PLA-801D cells was changed, and the antisense hTERT gene can act as an agent for specific inhibiting to p53 mRNA and the mutated p53 protein expression. Conclusion:The results suggest that full-length antisense hTERT can directed suppressed p53 mRNA expression level and down regulated mutated p53 protein.