Abstract:Objective:To construct the double-H1promoters siRNA expression cassettes (SECs) targeted to human telomere retrotranscriptase (hTERT), and investigate the interfering effect of its siRNA on hTERT gene expression in HepG2 cells.
Methods :SECs were constructed by fusing PCR, based on two different human telomerase hTERT gene fragments.When SEGs transferred into HepG2 cells respectively, the SECs were transcripted to the siRNA.The interfering effect of SECs on the telomerase activity in the cells was assessed by telomeric repeat amplification protocol (TRAP) and PCR-EIA.
Results:SEGs were successful constracted, and the telomerase activity was significantly inhibited when the HepG2 cells were tranfected with SECs.
Conclusions:The siRNA SECs display a definite RNA interference effect on the expression of telomerase. This method of SECs preparation can be applied for RNAi research in tumor inhibition.