The mechanism by which selleckchem hTERTp/CMV-dual-regulated TK expression can enhance the targeted killing of nasopharyngeal carcinoma cells need to be further investigated. In our previous study on hTERT-TK expression vector, the killing effect of TK under hTERT promoter, which is a much weaker than CMV promoter, is significantly reduced compared with that of TK under the non-selective promoter CMV. In consistence with our other reports [7–9], our results suggest that addition of CMV promoter can significantly enhance TK efficacy without changing its targeting controlled by hTERT. Wang [11, 12] proposed that
CMV can recognize specific binding sites of different activators, enhancers and promoters, therefore synergistically and dramatically promotes protein expression. In addition, co-effect of SV40 and CMV enhancers also enhance promoter activity because SV40 enhancer can effectively increase the amount of exogenous DNA in the nucleus. Therefore, the interference between hTERTp and CMV hindered the efficiency of vector. In this
study, we found that telomerase activities are significantly reduced in both NPC 5-8F and MCF-7 cells transfected with the enhanced vector after GCV treatment, but not changed in ECV cells transfected with the enhanced vector (Figure 4). One possible explanation is that the reduced telomerase activity in cells transfected with the enhanced vector is the result of the cell death induced by TK/GCV. We speculate that in the early stage of transfection of the enhanced vector, when GCV was not added into the cells, telomerase activity is temporally increased; selleck kinase inhibitor after adding GCV into the cells, cell numbers dramatically decreased resulting in the reduced telomerase activity. BAY 80-6946 However, we can not exclude other possibilities. Decreased telomerase activity has been shown to inhibit tumor proliferation. Transfection of eukaryotic vector containing antisense of hTERT in human gastric cancer SGC-7901 cells attenuated telomerase activity, reduced telomere length, decreased expressions of hTERT, bcL-2 and c-myC at mRNA and protein levels without changing hTR and
TP1 expression, inhibited cell proliferation and arrested the cells in G0/G1 phase . Injection of SGC-7901 cells Tyrosine-protein kinase BLK transfected with the eukaryotic vector containing antisense of hTERT did not induce tumor development in nude mice, whereas injection of control cells without transfection induced touchable tumor growth. Transfection of hTERT small interfering RNA had similar results . But it is more plausible that the mechanisms by which hTERT antisense or siRNA induced tumor apoptosis through reduced telomerase activity are different from that of the direct tumor killing of TK gene expression driven by hTERT promoter. To our knowledge, the effect of TK gene expression driven by CMV enhancer/hTERT promoter has not been previously studied in NPC.