PDGF-A promoter and enhancer elements provide efficient and selective antineoplastic gene therapy in multiple cancer types.
PDGF-A promoter and enhancer elements provide efficient and selective antineoplastic gene therapy in multiple cancer types.
复制标题
DOI：
10.1038/cgt.2008.92
复制
发表时间：
2009-04
影响因子：
6.4
通讯作者：
中科院分区：
文献类型：
--
作者：
由于缺乏具有有效的癌细胞特异性活性的转录控制元件，因此抑制基因疗法的发展受到损害。我们研究了血小板衍生生长因子-A（PDGF-A）基因启动子（AChP）和5 '末端增强子（ACE 66）元件的效用，这些元件在许多人类癌症中过度活跃。在多种肿瘤细胞系中测试这些元素的功效，包括在细胞培养物中以及作为无胸腺裸鼠中的肿瘤外植体。质粒和病毒载体的构建与乙酰胆碱酯酶启动子单独或与三个拷贝的ACE 66增强子融合的原型自杀基因，胸苷激酶（TK）的表达。ACE/AChP和AChP盒在多种肿瘤细胞系中引起更昔洛韦（GCV）诱导的细胞毒性。ACE增强子元件还表现出与胎盘和肝脏特异性启动子元件的协同作用。含有AChP-TK盒的腺病毒在培养的肿瘤细胞系中产生GCV敏感性的显著增加，以及GCV诱导的体内U87 MG胶质母细胞瘤外植体的消退。TK表达分布在接受治疗性病毒的肿瘤中，而TdT介导的dUTP缺口末端标记分析显示许多区域发生凋亡。血管化和网状纤维网络在病毒GCV治疗的肿瘤中不太明显，表明原发性和基质细胞类型都可能被靶向。这些研究为PDGF-A启动子和ACE 66增强子在多种人类癌症的PDGF-A基因治疗中的应用提供了原理证明。
Development of antineoplastic gene therapies is impaired by a paucity of transcription control elements with efficient, cancer cell-specific activity. We investigated the utility of promoter (AChP) and 5’-distal enhancer (ACE66) elements from the platelet-derived growth factor-A (PDGF-A) gene, which are hyperactive in many human cancers. Efficacy of these elements was tested in multiple tumor cell lines, both in cell culture and as tumor explants in athymic nude mice. Plasmid and viral vectors were constructed with the AChP promoter alone or in fusion with three copies of the ACE66 enhancer for expression of the prototype suicide gene, thymidine kinase (TK). ACE/AChP and AChP cassettes elicited ganciclovir (GCV)-induced cytotoxicity in multiple tumor cell lines. The ACE enhancer element also exhibited synergism with placental and liver-specific promoter elements. An adenovirus containing the AChP-TK cassette produced striking increases in GCV sensitivity in cultured tumor cell lines, as well as GCV-induced regression of U87 MG glioblastoma explants in vivo. TK expression was distributed throughout tumors receiving the therapeutic virus, whereas TdT-mediated dUTP nick end labeling analysis revealed numerous regions undergoing apoptosis. Vascularization and reticulin fiber networks were less pronounced in virus-GCV-treated tumors, suggesting that both primary and stromal cell types may have been targeted. These studies provide proof-of-principle for utility of the PDGF-A promoter and ACE66 enhancer in antineoplastic gene therapy for a diverse group of human cancers.
登录
查看更多内容
影响因子：
64.8
作者：
BETSHOLTZ, C;JOHNSSON, A;SCOTT, J
通讯作者：
SCOTT, J
影响因子：
4.8
作者：
HECKERT, LL;SCHULTZ, K;NILSON, JH
通讯作者：
NILSON, JH
影响因子：
4.1
作者：
KAETZEL, DM;MAUL, RS;COYNE, DW
通讯作者：
COYNE, DW
影响因子：
5.3
作者：
Long, LY;Spear, BT
通讯作者：
Spear, BT
影响因子：
20.1
作者：
Khachigian, LM;Santiago, FS;Johnson, AC
通讯作者：
Johnson, AC