Seth P, Brinkmann U, Schwartz G N, Katayose D, Gress R, Cowan K
Medicine Branch, National Cancer Institute, NIH, Bethesda, Maryland 20892, USA.
Cancer Res. 1996 Mar 15;56(6):1346-51.
To examine the potential use of adenovirus vectors in cancer gene therapy as a mechanism for purging bone marrow cells of possible breast cancer contaminants, we compared the infection efficiency of adenovirus and the transfection efficiency of plasmid DNA in the presence of adenovirus in human breast cancer and bone marrow cells. Following infection of breast cancer cells with an adenovirus expressing beta-galactosidase gene, high levels of beta-galactoside activity were observed. No beta-galactosidase activity was observed in low-density human bone marrow cells. A replication-deficient adenovirus mutant dl312 enhanced the transfection efficiency of a plasmid DNA-expressing beta-galactosidase gene into breast cancer cells, and addition of a liposome, lipofectamine, further enhanced the transfection efficiency. In contrast, human bone marrow cells treated under the same conditions expressed very low levels of transfected beta-galactosidase DNA. Transfection of cells with plasmid DNA expressing a truncated but fully active Pseudomonas exotoxin gene in the presence of dl312 and lipofectamine resulted in marked breast cancer cell killing, whereas colony-forming unit granulocyte-macrophage (CFU-GM) were relatively resistant to these treatments. A recombinant adenovirus expressing human wild-type p53 protein (AdWTp53) was also highly cytotoxic to breast tumor cells. Infection of breast cancer cells with AdWTp53 (100 plaque-forming units/cell) resulted in 100% loss of the clonogenicity of breast tumor cells. However, colony formation from CFU-GM was relatively resistant to the cytotoxic effects of AdWTp53 alone or in the presence of pULI100 plasmid and lipofectamine. On the basis of these results, it is proposed that human adenoviruses are potentially useful for cancer gene therapy and bone marrow purging.
为了研究腺病毒载体在癌症基因治疗中作为清除骨髓细胞中可能存在的乳腺癌污染物的一种机制的潜在用途，我们比较了腺病毒在人乳腺癌细胞和骨髓细胞中的感染效率以及腺病毒存在下质粒DNA的转染效率。用表达β-半乳糖苷酶基因的腺病毒感染乳腺癌细胞后，观察到高水平的β-半乳糖苷活性。在低密度人骨髓细胞中未观察到β-半乳糖苷酶活性。一种复制缺陷型腺病毒突变体dl312提高了表达β-半乳糖苷酶基因的质粒DNA转入乳腺癌细胞的转染效率，添加脂质体Lipofectamine进一步提高了转染效率。相比之下，在相同条件下处理的人骨髓细胞表达的转染β-半乳糖苷酶DNA水平非常低。在dl312和Lipofectamine存在下，用表达截短但具有完全活性的铜绿假单胞菌外毒素基因的质粒DNA转染细胞导致显著的乳腺癌细胞杀伤，而集落形成单位粒细胞-巨噬细胞（CFU-GM）对这些处理相对耐药。一种表达人野生型p53蛋白的重组腺病毒（AdWTp53）对乳腺肿瘤细胞也具有高度细胞毒性。用AdWTp53（100个噬斑形成单位/细胞）感染乳腺癌细胞导致乳腺肿瘤细胞的克隆形成能力100%丧失。然而，CFU-GM的集落形成对单独的AdWTp53或在存在pULI100质粒和Lipofectamine的情况下的细胞毒性作用相对耐药。基于这些结果，有人提出人腺病毒在癌症基因治疗和骨髓净化方面具有潜在用途。
