Eastham J A, Hall S J, Sehgal I, Wang J, Timme T L, Yang G, Connell-Crowley L, Elledge S J, Zhang W W, Harper J W
Scott Department of Urology, Baylor College of Medicine, Houston, Texas 77030, USA.
Cancer Res. 1995 Nov 15;55(22):5151-5.
We introduced the gene for wild-type human p53 or p21, a critical downstream mediator of p53-induced growth suppression, into a p53-deficient mouse prostate cancer cell line using a recombinant adenoviral vector (Ad5CMV-p53 or Ad5CMV-p21). Elevated levels of endogenous mouse p21 mRNA provided evidence for the functional activity of virally transduced p53. Functional activity of viral-transduced p21 was demonstrated through immunoprecipitation of cellular protein extracts, which showed that the viral-transduced p21 associates with cyclin-dependent kinase 2 and was sufficient to down-regulate the activity of the cyclin-dependent kinase by approximately 65%. In vitro growth assays revealed significantly higher growth suppression after Ad5CMV-p21 infection compared to Ad5CMV-p53. In vivo studies in syngeneic male mice with established s.c. prostate tumors demonstrated that the rate of growth and final tumor volume were reduced to a much greater extent in mice that received intratumor injection of Ad5CMV-p21 compared to Ad5CMV-p53. In addition, the survival of host animals bearing tumors that were infected with Ad5CMV-p21, but not Ad5CMV-p53, was significantly extended. These data suggest that Ad5CMV-p21 may be effective as a therapeutic agent for prostate cancer.
我们使用重组腺病毒载体（Ad5CMV-p53 或 Ad5CMV-p21），将野生型人类 p53 基因或 p21（p53 诱导生长抑制的关键下游介质）导入 p53 缺陷的小鼠前列腺癌细胞系。内源性小鼠 p21 mRNA 水平升高为病毒转导的 p53 的功能活性提供了证据。通过对细胞蛋白提取物进行免疫沉淀，证明了病毒转导的 p21 的功能活性，结果显示病毒转导的 p21 与细胞周期蛋白依赖性激酶 2 结合，并且足以将细胞周期蛋白依赖性激酶的活性下调约 65%。体外生长试验显示，与 Ad5CMV-p53 相比，Ad5CMV-p21 感染后生长抑制作用显著增强。在具有皮下前列腺肿瘤的同基因雄性小鼠中进行的体内研究表明，与 Ad5CMV-p53 相比，接受瘤内注射 Ad5CMV-p21 的小鼠的生长速率和最终肿瘤体积降低的程度更大。此外，携带感染 Ad5CMV-p21 而非 Ad5CMV-p53 的肿瘤的宿主动物的存活时间显著延长。这些数据表明，Ad5CMV-p21 可能作为前列腺癌的治疗剂有效。
