Watanabe Masami, Sakaguchi Masakiyo, Kinoshita Rie, Kaku Haruki, Ariyoshi Yuichi, Ueki Hideo, Tanimoto Ryuta, Ebara Shin, Ochiai Kazuhiko, Futami Junichiro, Li Shun-Ai, Huang Peng, Nasu Yasutomo, Huh Nam-Ho, Kumon Hiromi
Center for Innovative Clinical Medicine, Okayama University Hospital, Okayama, Japan.
Department of Cell Biology, Okayama University, Okayama, Japan.
Oncol Rep. 2014 Mar;31(3):1089-95. doi: 10.3892/or.2013.2958. Epub 2013 Dec 31.
Gene expression systems with various promoters, including the cytomegalovirus (CMV) promoter, have been developed to increase the gene expression in a variety of normal and cancer cells. In particular, in the clinical trials of cancer gene therapy, a more efficient and robust gene expression system is required to achieve sufficient therapeutic outcomes. By inserting the triple translational enhancer sequences of human telomerase reverse transcriptase (hTERT), Simian virus 40 (SV40) and CMV downstream of the sequence of the BGH polyA, we were able to develop a novel gene expression system that significantly enhances the expression of the genes of interest. We termed this novel gene expression cassette the super gene expression (SGE) system, and herein verify the utility of the SGE cassette for a replication-deficient adenoviral vector. We newly developed an adenoviral vector expressing the tumor suppressor, reduced expression in immortalized cells (REIC)/Dickkopf-3 (Dkk-3), based on the CMV promoter-driven SGE system (Ad-SGE-REIC) and compared the therapeutic utility of Ad-SGE-REIC with that of the conventional adenoviral vectors (Ad-CMV-REIC or Ad-CAG-REIC). The results demonstrated that the CMV promoter-SGE system allows for more potent gene expression, and that the Ad-SGE-REIC is superior to conventional adenoviral systems in terms of the REIC protein expression and therapeutic effects. Since the SGE cassette can be applied for the expression of various therapeutic genes using various vector systems, we believe that this novel system will become an innovative tool in the field of gene expression and gene therapy.
已经开发出具有各种启动子的基因表达系统，包括巨细胞病毒（CMV）启动子，以增加多种正常细胞和癌细胞中的基因表达。特别是在癌症基因治疗的临床试验中，需要更高效、强大的基因表达系统来实现足够的治疗效果。通过在BGH polyA序列下游插入人端粒酶逆转录酶（hTERT）、猿猴病毒40（SV40）和CMV的三联翻译增强子序列，我们能够开发出一种新型基因表达系统，该系统可显著增强目的基因的表达。我们将这个新型基因表达盒称为超基因表达（SGE）系统，并在此验证SGE盒在复制缺陷型腺病毒载体中的实用性。我们基于CMV启动子驱动的SGE系统（Ad-SGE-REIC）新开发了一种表达肿瘤抑制因子、永生化细胞中表达降低（REIC）/Dickkopf-3（Dkk-3）的腺病毒载体，并将Ad-SGE-REIC与传统腺病毒载体（Ad-CMV-REIC或Ad-CAG-REIC）的治疗效用进行了比较。结果表明，CMV启动子-SGE系统可实现更强的基因表达，并且Ad-SGE-REIC在REIC蛋白表达和治疗效果方面优于传统腺病毒系统。由于SGE盒可用于使用各种载体系统表达各种治疗性基因，我们相信这个新系统将成为基因表达和基因治疗领域的创新工具。
