Von Gruenigen V. E., O'Boyle J. D., Coleman R. L., Wilson D., Miller D. S., Mathis J. M.
Division of Gynecologic Oncology, Departments of Obstetrics and Gynecology and Biochemistry, Hammon Center for Therapeutic Oncologic Research, The University of Texas Southwestern Medical Center, Dallas, Texas, and Introgen Therapeutics, Inc., Houston, Texas USA.
Int J Gynecol Cancer. 1999 Sep;9(5):365-372. doi: 10.1046/j.1525-1438.1999.99040.x.
The purpose of this study was to determine the efficacy of adenovirus-based p53 gene therapy in the treatment of ovarian cancer using an intraperitoneal microscopic tumor animal model system. Adenovirus-mediated wild-type p53 gene was introduced into the NIH:OVCAR-3 human ovarian cancer cell line in vitro and in vivo. In order to study microscopic intraperitoneal tumor, athymic nude mice were inoculated intraperitoneally (i.p.) with 1 x 107 OVCAR-3 cells and observed for tumor growth. Three days after inoculation with OVCAR-3 cells, the mice were divided into 3 treatment groups. One group received three daily i.p. injections of 1 x 108 pfu Ad-CMV-p53, a second group received three daily i.p. injection of 1 x 108 pfu of the control adenovirus construct expressing beta galactosidase (Ad-CMV-betagal) and a third group received three daily i.p. injections of normal saline. Adenovirus-mediated introduction of the wild-type p53 gene in the ovarian cancer cell line resulted in transient high levels of p53 protein for 24-48 h. Cell cycle analysis revealed G1 arrest, as well as the appearance of apoptosis. In vitro cell growth assays showed growth inhibition of cancer cells infected with Ad-CMV-p53 compared to cells infected with Ad-CMV-betagal or normal saline. There was a significant increase in survival in the Ad-CMV-p53 adenovirus treated animals compared to the PBS treated animals (P = 0.004). Likewise, the survival in Ad-CMV-p53 treated mice was also significantly greater than mice treated with Ad-CMV-betagal (P < 0.0001). These results demonstrated that Ad-CMV-p53 treatment is effective in inhibiting tumor growth and prolonging survival in this microscopic cancer xenograft model. The results of this study constitute a step in translating promising in vitro and in vivo data from an adenovirus-based gene therapeutic model system into practical and scientifically based human cancer therapeutic trials.
本研究的目的是利用腹腔内微小肿瘤动物模型系统，确定基于腺病毒的p53基因疗法治疗卵巢癌的疗效。腺病毒介导的野生型p53基因在体外和体内被导入NIH:OVCAR-3人卵巢癌细胞系。为了研究腹腔内微小肿瘤，将无胸腺裸鼠经腹腔接种1×10⁷个OVCAR-3细胞，并观察肿瘤生长情况。接种OVCAR-3细胞三天后，将小鼠分为3个治疗组。一组每天经腹腔注射1×10⁸ pfu的Ad-CMV-p53，第二组每天经腹腔注射1×10⁸ pfu表达β-半乳糖苷酶的对照腺病毒构建体（Ad-CMV-βgal），第三组每天经腹腔注射生理盐水。腺病毒介导的野生型p53基因导入卵巢癌细胞系导致p53蛋白在24至48小时内短暂高水平表达。细胞周期分析显示G1期阻滞以及凋亡的出现。体外细胞生长试验表明，与感染Ad-CMV-βgal或生理盐水的细胞相比，感染Ad-CMV-p53的癌细胞生长受到抑制。与经PBS处理的动物相比，经Ad-CMV-p53腺病毒处理的动物存活率显著提高（P = 0.004）。同样，经Ad-CMV-p53处理的小鼠存活率也显著高于经Ad-CMV-βgal处理的小鼠（P < 0.0001）。这些结果表明，在这个微小癌异种移植模型中，Ad-CMV-p53治疗可有效抑制肿瘤生长并延长生存期。本研究结果是将基于腺病毒的基因治疗模型系统中体外和体内的有前景数据转化为实际的、基于科学的人类癌症治疗试验的重要一步。
