Turturro F, Heineke H L, Drevyanko T F, Link C J, Seth P
Human Gene Therapy Research Institute, John Stoddard Cancer Center, Des Moines, IA, USA.
Gene Ther. 2000 Jun;7(11):930-3. doi: 10.1038/sj.gt.3301186.
Adenovirus-p53-mediated apoptosis has been extensively evaluated in animal xenografts derived from human epithelial tumors and recently began testing in phase I clinical trials, but has not been evaluated for lymphoid malignancies. Cell lines derived from anaplastic large cell lymphoma (ALCL) carrying the t(2;5) translocation are efficiently transduced by adenoviral vector expressing p53 and undergo apoptosis. To test the in vivo efficiency of adenovirus-mediated-p53 expression and apoptosis induction, SUDHL-1 cells (derived from human ALCL) were injected subcutaneously into athymic nude mice. Cells from the xenograft had typical morphology of human ALCL by standard hematoxylin-eosin staining, CD5+, CD45+ and CD30+ immunophenotype, the t(2;5) translocation by PCR. Six tumors from an initial set of mice were evaluated for apoptosis by TUNEL and for necrosis by hematoxylin-eosin staining 48-72 h after injection with 1 x 108 p.f.u. of AdWTp53 (adenoviral vector expressing p53), of AdNull (adenoviral vector backbone) and PBS (mock), respectively. TUNEL staining was positive only in tumors injected with AdWTp53 and was mainly localized around the needle track. Differences of the means of the counts of the necrotic cells were statistically significant at P = 0.02 between AdWTp53 and mock and only borderline between AdWTp53 and AdNull. Twenty-three tumors from a separate set of mice were subsequently injected with AdWTp53, AdNull and PBS and evaluated for in vivo tumor response. Three total injections of viral vectors (1 x 108 p.f.u.) and PBS were given every 48-72 h. Only tumors injected with AdWTp53 showed tumor growth inhibition with a mean final tumor volume that was statistically significantly smaller than AdNull (P = 0.007) and mock (P = 0.002). Based on these results we foresee a potential application of adenovirus-mediated p53 apoptosis as gene therapy of lymphomas.
腺病毒介导的p53凋亡已在源自人类上皮肿瘤的动物异种移植模型中得到广泛评估，最近已开始进行I期临床试验，但尚未对淋巴系统恶性肿瘤进行评估。携带t(2;5)易位的间变性大细胞淋巴瘤（ALCL）来源的细胞系能被表达p53的腺病毒载体有效转导并发生凋亡。为了测试腺病毒介导的p53表达及凋亡诱导在体内的效率，将SUDHL-1细胞（源自人类ALCL）皮下注射到无胸腺裸鼠体内。通过标准苏木精-伊红染色，移植瘤细胞具有人类ALCL的典型形态，通过免疫表型分析显示为CD5+、CD45+和CD30+，通过PCR检测存在t(2;5)易位。在分别注射1×108 p.f.u.的AdWTp53（表达p53的腺病毒载体）、AdNull（腺病毒载体骨架）和PBS（假注射）48 - 72小时后，对最初一组小鼠的6个肿瘤进行TUNEL凋亡检测和苏木精-伊红染色坏死检测。TUNEL染色仅在注射AdWTp53的肿瘤中呈阳性，且主要位于针道周围。AdWTp53与假注射组之间坏死细胞计数平均值的差异在P = 0.02时有统计学意义，而AdWTp53与AdNull组之间仅为临界差异。随后对另一组小鼠的23个肿瘤分别注射AdWTp53、AdNull和PBS，并评估体内肿瘤反应。每48 - 72小时进行三次病毒载体（1×108 p.f.u.）和PBS的注射。只有注射AdWTp53的肿瘤显示出肿瘤生长抑制，其最终平均肿瘤体积在统计学上显著小于AdNull（P = 0.007）和假注射组（P = 0.002）。基于这些结果，我们预见腺病毒介导的p53凋亡作为淋巴瘤基因治疗具有潜在应用价值。
