Badie B, Hunt K, Economou J S, Black K L
Division of Neurosurgery, University of California Los Angeles School of Medicine.
Neurosurgery. 1994 Nov;35(5):910-5; discussion 915-6. doi: 10.1227/00006123-199411000-00016.
The dismal results of conventional therapy for primary malignant brain tumors has justified exploring gene therapy approaches for this disease. Transduction of animal brain tumor models in vivo has been reported previously with retroviruses and herpes viruses. Because adenoviruses have the advantage of transducing quiescent and actively dividing tumor cells, they may prove to be more effective in such therapy. We used a replication-deficient recombinant adenovirus bearing the Escherichia coli beta-galactosidase gene in a rat C6 glioma tumor model. Transduced cells were detected by X-5-bromo-4-chloro-3-indolyl beta-D-galactoside staining to reveal beta-galactosidase activity. Initial experiments in vitro showed 50% and 90% transduction at vector titers of approximately 10(7) and 10(8) plaque-forming units/ml, respectively. Although no cytopathic effects were seen at 10(7) plaque-forming units/ml, more than 50% reduction in tumor cell growth was noted at 10(8) plaque-forming units/ml both in vitro and in vivo. Stereotactic delivery of the recombinant adenovirus into the frontal lobe of normal rat brains resulted in intense staining of all cell types, that is, neurons, astrocytes, and ependymal cells. Stereotactic injection into C6 glioma brain tumors in rats stained 25 to 30% of the tumor cells. We conclude that adenovirus vectors can be used to transfer genes to central nervous system tumors in vivo. Using stereotactic delivery, adenovirus vectors can transfer genes into the central nervous system intended for tumor therapy.
原发性恶性脑肿瘤传统治疗方法的效果不佳，这使得探索针对该疾病的基因治疗方法具有合理性。此前已有报道使用逆转录病毒和疱疹病毒在体内转导动物脑肿瘤模型。由于腺病毒具有转导静止和活跃分裂肿瘤细胞的优势，它们可能在这种治疗中被证明更有效。我们在大鼠C6胶质瘤肿瘤模型中使用了携带大肠杆菌β-半乳糖苷酶基因的复制缺陷型重组腺病毒。通过X-5-溴-4-氯-3-吲哚基β-D-半乳糖苷染色检测转导细胞，以揭示β-半乳糖苷酶活性。体外初步实验显示，在载体滴度约为10(7)和10(8) 噬斑形成单位/毫升时，转导率分别为50%和90%。虽然在10(7) 噬斑形成单位/毫升时未观察到细胞病变效应，但在10(8) 噬斑形成单位/毫升时，体外和体内肿瘤细胞生长均减少了50%以上。将重组腺病毒立体定向递送至正常大鼠脑额叶导致所有细胞类型，即神经元、星形胶质细胞和室管膜细胞出现强烈染色。立体定向注射到大鼠C6胶质瘤脑肿瘤中，使25%至30%的肿瘤细胞染色。我们得出结论，腺病毒载体可用于在体内将基因转移至中枢神经系统肿瘤。使用立体定向递送，腺病毒载体可将基因转移至中枢神经系统用于肿瘤治疗。
