Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry , Russian Academy of Sciences, Miklukho-Maklaya St., 16/10, Moscow, Russia, 117997.
Acta Naturae. 2013 Jul;5(3):79-83.
Core promoters with adjacent regions of the human genes CDC6, POLD1, CKS1B, MCM2, and PLK1 were cloned into a pGL3 vector in front of the Photinus pyrails gene Luc in order to study the tumor specificity of the promoters. The cloned promoters were compared in their ability to direct luciferase expression in different human cancer cells and in normal fibroblasts. The cancer-specific promoter BIRC5 and non-specific CMV immediately early gene promoter were used for comparison. All cloned promoters were shown to be substantially more active in cancer cells than in fibroblasts, while the PLK1 promoter was the most cancer-specific and promising one. The specificity of the promoters to cancer cells descended in the series PLK1, CKS1B, POLD1, MCM2, and CDC6. The bidirectional activity of the cloned CKS1B promoter was demonstrated. It apparently directs the expression of the SHC1 gene, which is located in a "head-to-head" position to the CKS1B gene in the human genome. This feature should be taken into account in future use of the CKS1B promoter. The cloned promoters may be used in artificial genetic constructions for cancer gene therapy.
为了研究启动子的肿瘤特异性，将人类基因 CDC6、POLD1、CKS1B、MCM2 和 PLK1 的核心启动子及其邻近区域克隆到 Photinus pyrails 基因 Luc 的 pGL3 载体前面。比较了克隆启动子在不同人癌细胞和正常成纤维细胞中指导荧光素酶表达的能力。使用已克隆的肿瘤特异性 BIRC5 启动子和非特异性 CMV 即刻早期基因启动子进行比较。所有克隆的启动子在癌细胞中的活性都明显高于成纤维细胞，而 PLK1 启动子是最具肿瘤特异性和最有前途的启动子。启动子对癌细胞的特异性在 PLK1、CKS1B、POLD1、MCM2 和 CDC6 这一系列中逐渐降低。证明了克隆的 CKS1B 启动子具有双向活性。它显然指导了位于人类基因组中与 CKS1B 基因“头对头”位置的 SHC1 基因的表达。在未来使用 CKS1B 启动子时，应考虑到这一特征。克隆的启动子可用于癌症基因治疗的人工遗传构建。
