Teoh G, Chen L, Urashima M, Tai Y T, Celi L A, Chen D, Chauhan D, Ogata A, Finberg R W, Webb I J, Kufe D W, Anderson K C
Department of Adult Oncology, Dana-Farber Cancer Institute, Boston, MA; and the Department of Haematology, Singapore General Hospital, Singapore.
Blood. 1998 Dec 15;92(12):4591-601.
Adenoviruses are efficient gene delivery agents for a variety of neoplasms. In the present study, we have investigated the use of adenoviruses for the delivery of the thymidine kinase (tk) gene into multiple myeloma (MM) cells. We first demonstrated that MM cell lines and MM patient cells express both adenovirus receptors as well as the DF3/MUC1 protein, thus providing a rationale for using adenoviruses to selectively deliver genes under the control of the DF3 promoter. By using an adenoviral construct containing beta-galactosidase (beta-gal) gene driven by the DF3 promoter (Ad. DF3-betagal), we demonstrate greater than 80% transduction efficiency in OCI-My5 and RPMI 8226 MM cell lines at a multiplicity of infection of 1 to 100. Importantly, transduction with the tk gene driven by the DF3 promoter (Ad.DF3-tk) followed by treatment with 50 micromol/L ganciclovir (GCV) purged >/=6 log of contaminating OCI-My5 and RPMI 8226 MM cells within bone marrow mononuclear cells. In contrast, normal human hematopoietic progenitor cell number was unaffected under these conditions. Selectivity of DF3/MUC1 promoter was further confirmed, because Ad.DF3-betagal or Ad.DF3-tk did not transduce MUC1-negative HeLa cervical carcinoma cells. In addition, GCV treatment of Ad.DF3-tk-transduced RPMI 8226 MM cells did not induce a significant bystander effect. These findings demonstrate that transduction with Ad vectors using a tumor-selective promoter provides a highly efficient and selective approach for the ex vivo purging of MM cells.
腺病毒是用于多种肿瘤的高效基因传递载体。在本研究中，我们研究了利用腺病毒将胸苷激酶（tk）基因传递到多发性骨髓瘤（MM）细胞中的情况。我们首先证明MM细胞系和MM患者细胞同时表达腺病毒受体以及DF3/MUC1蛋白，从而为利用腺病毒在DF3启动子控制下选择性传递基因提供了理论依据。通过使用一种含有由DF3启动子驱动的β-半乳糖苷酶（β-gal）基因的腺病毒构建体（Ad.DF3-βgal），我们发现在感染复数为1至100时，OCI-My5和RPMI 8226 MM细胞系中的转导效率超过80%。重要的是，用DF3启动子驱动的tk基因（Ad.DF3-tk）进行转导，随后用50 μmol/L更昔洛韦（GCV）处理，可清除骨髓单个核细胞中≥6 log的污染OCI-My5和RPMI 8226 MM细胞。相比之下，在这些条件下正常人造血祖细胞数量未受影响。DF3/MUC1启动子的选择性得到进一步证实，因为Ad.DF3-βgal或Ad.DF3-tk不能转导MUC1阴性的HeLa宫颈癌细胞。此外，用GCV处理Ad.DF3-tk转导的RPMI 8226 MM细胞未诱导明显的旁观者效应。这些发现表明，使用肿瘤选择性启动子的腺病毒载体转导为体外清除MM细胞提供了一种高效且选择性的方法。
