Hamada K, Zhang W W, Alemany R, Wolf J, Roth J A, Mitchell M F
Department of Thoracic and Cardiovascular Surgery, University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Gynecol Oncol. 1996 Mar;60(3):373-9. doi: 10.1006/gyno.1996.0057.
Human papillomavirus (HPV) has been identified in the majority of invasive cancers of the uterine cervix sampled and has been found to contribute in a significant way to the genesis of human cervical cancer. HPV has two transforming genes that encode the oncoproteins E6 and E7. E6 can form complexes with p53 and promote p53 degradation. We introduced wild-type p53 into a cervical cancer cell line via a recombinant adenoviral vector, Ad5CMV-p53. Human cervical cancer cell line HeLa, which has HPV type 18 and wild-type p53, was used in this study. Cells were grown in RPMI medium supplemented with 10% heat-inactivated fetal bovine serum. Ad5CMV-p53 was created by inserting the cytomegalovirus promoter, wild-type p53 cDNA, and SV40 polyadenylation signal in a minigene cassette into the E1-deleted region of the modified Ad5 adenovirus. The transduction efficiency was 100% when a dose ensuring a multiplicity of infection of 100 or greater was used. The p53 protein was detected in Ad5CMV-p53-infected cells by immunohistochemical and Western blot analyses. The growth of the Ad5CMV-p53-infected cells was greatly suppressed as detected by both cell count and [3H]thymidine incorporation assay. These data suggest that transfection of HPV-positive cervical cancer cells with a wild-type p53 gene in a form such as Ad5CMV-p53 is a potential novel therapy for cervical cancer.
在大多数采样的子宫颈浸润癌中已鉴定出人乳头瘤病毒（HPV），并且发现其在人类宫颈癌的发生中起重要作用。HPV有两个转化基因，编码癌蛋白E6和E7。E6可与p53形成复合物并促进p53降解。我们通过重组腺病毒载体Ad5CMV-p53将野生型p53导入宫颈癌细胞系。本研究使用了具有18型HPV和野生型p53的人宫颈癌细胞系HeLa。细胞在补充有10%热灭活胎牛血清的RPMI培养基中培养。Ad5CMV-p53是通过将巨细胞病毒启动子、野生型p53 cDNA和SV40聚腺苷酸化信号插入小基因盒中，并导入修饰的Ad5腺病毒的E1缺失区域而构建的。当使用确保感染复数为100或更高的剂量时，转导效率为100%。通过免疫组织化学和蛋白质印迹分析在Ad5CMV-p53感染的细胞中检测到p53蛋白。通过细胞计数和[3H]胸苷掺入试验检测发现，Ad5CMV-p53感染的细胞生长受到极大抑制。这些数据表明，用Ad5CMV-p53等形式的野生型p53基因转染HPV阳性宫颈癌细胞是一种潜在的宫颈癌新疗法。
