Clone of the hTERT Promoter and Its Specific Transcriptional Activity in Human Gastric Caner Cells

Objective:To clone a human telomerase reverse transcriptase(hTERT)core promoter,and to investigate the tran- scriptional activity of the hTERT promoter in the human gastric cancer cells SGC7901 and the normal human fibroblast cells HLF.Meth- ods:The hTERT core promoter was amplified by PCR using the total genomic DNA of Hela cells as sample DNA and then cloned into pGL3-Basic vector.The recombinant was named hTERT-pGL3Basic.This vector was transfected into SGC7901 cells with high telom- erase activity and into RLF ceils without telomerase activity to detect the transcriptional activities of the hTERT promoter by cationic li- posome.Results:The hTERT core promoter was amplified by PCR successfully.The results of restriction enzyme digestion and PCR in- dicated that the hTERT-pGL3Basic expression vector was constructed successfully as the design.Considering transcriptional activity of pGL3-control in each cell line as 100%,luciferase assay showed the relative transcriptional activity ofhTERT promoter in SGC7901 was 21.5% and that of hTERT promoter in HLF cells was only 0.40%.Conclusions:The hTERT promoter has tumor specificity and may be useful in targeting gene therapy for tumor.