癌胚抗原的转录调控序列:识别和利用肿瘤特异性的胞嘧啶脱氨酶基因治疗。
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理查兹CA,奥斯汀EA,休伯
癌胚抗原的转录调控序列:识别和利用肿瘤特异性的胞嘧啶脱氨酶基因治疗。
嗡嗡声其他基因。1995年7月,6 (7):881 - 93。
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7578407 (在PubMed]
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5 '人类癌胚抗原(CEA)基因序列分析了用荧光素酶报告基因检测。这种分析确定所需重要cis-acting序列选择性表达CEA-positive细胞。超过50东航/荧光素酶记者克隆构建和分析在两个CEA-positive和两个CEA-negative细胞系。CEA的序列分析扩展转化开始14.5 kb CEA基因的5 '。从东航408 - bp地区也检查了3 '端非翻译区对记者基因活性的影响。CEA启动子位于基地之间的-90和69 +的转录启动站点。序列之间的-41年和-18年从CEA启动子表达的必要条件。Multimerization序列之间的-89年和-40年导致复制number-related增加对CEA-positive细胞表达水平和选择性。两个东航的上游地区,-13.6到-10.7 kb或-6.1到-4.0 kb,与独处时启动子导致高层,选择性CEA-positive细胞系中表达。几个东航/荧光素酶结构显示80 - 120倍高表达CEA-positive细胞系表达相比CEA-negative Hep3B细胞。 The expression from these constructs was quite strong in CEA-positive cells, being two- to four-fold higher than an SV40 enhancer/promoter construct. The most promising CEA transcriptional regulatory sequences were used to regulate the expression of cytosine deaminase (CD) in stable cell lines. The expression of CD was assessed directly by an enzymatic assay and indirectly by determining the in vitro IC50 to 5-fluorocytosine (5FC). The chimeric gene pCEA/CD-145 displayed the desired expression spectrum--high-level expression in the CEA-positive cells and low-level expression in CEA-negative cells. CD expression from this chimera correlated well with the expression of the endogenous CEA gene. Treatment of mice bearing NCI H508 pCEA/CD-145 tumor xenografts with 5FC lead to significant antitumor effects in vivo. The CEA/CD chimeric gene should be useful for tumor-specific suicide gene therapy of CEA-positive tumors.