Kim Sung Jin, Lee Han Saem, Shin June Ho, Kim Chul Geun, Jeong Sunjoo, Park Keerang, Choe Han, Lee Heuiran
Department of Microbiology, University of Ulsan College of Medicine, 138-736 Seoul, Korea.
Oncol Rep. 2006 Nov;16(5):975-9.
Although the human telomerase reverse transcriptase (hTERT) promoter can regulate cancer-specific genes, it is generally too weak to be effective. We therefore attempted to improve the potency of synthetic hTERT promoters by fusing the core element (E) of the hTERT promoter (H) and the tripartite leader sequence (T) from human adenovirus 5 in a combinatorial manner. To determine the potential as cancer-specific promoters, we measured luciferase activity driven by the chimeric hTERT promoters in human cancer cells. Among various constructs, the E3-H-T promoter induced the strongest luciferase activity in all the tested cancer cells. SK-Hep1 and Hela cells experienced 1000- and 11-fold higher expression than the basic hTERT promoter, respectively. Relative to the SV40 universal promoter, the E3-H-T promoter led to higher levels of gene expression. Using EMSA, we found that the hTERT enhancer region was specifically bound to c-Myc and Sp1. Thus, the data suggest that the E3-H-T promoter with up-regulated cancer-specific gene expression could be useful in cancer gene therapy.
尽管人端粒酶逆转录酶（hTERT）启动子可调控癌症特异性基因，但它通常过于微弱而无法有效发挥作用。因此，我们尝试通过将hTERT启动子的核心元件（E）与来自人腺病毒5的三联前导序列（T）以组合方式融合，来提高合成hTERT启动子的效力。为了确定作为癌症特异性启动子的潜力，我们测量了嵌合hTERT启动子在人癌细胞中驱动的荧光素酶活性。在各种构建体中，E3-H-T启动子在所有测试的癌细胞中诱导出最强的荧光素酶活性。SK-Hep1细胞和Hela细胞的表达分别比基本hTERT启动子高1000倍和11倍。相对于SV40通用启动子，E3-H-T启动子导致更高水平的基因表达。使用电泳迁移率变动分析（EMSA），我们发现hTERT增强子区域与c-Myc和Sp1特异性结合。因此，数据表明具有上调的癌症特异性基因表达的E3-H-T启动子可能在癌症基因治疗中有用。
