Growth inhibition of human cervical cancer cells with the recombinant adenovirus p53 in vitro.
Growth inhibition of human cervical cancer cells with the recombinant adenovirus p53 in vitro.
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体外重组腺病毒p53对人宫颈癌细胞生长的抑制作用。
DOI：
10.1006/gyno.1996.0057
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发表时间：
1996
影响因子：
4.7
通讯作者：
Mitchell,MF
中科院分区：
文献类型：
--
作者：
Hamada,K;Zhang,WW;Alemany,R;Wolf,J;Roth,JA;Mitchell,MF
人乳头瘤病毒（HPV）已被确定在大多数的子宫颈的浸润性癌采样，并已发现在一个显着的方式有助于人类宫颈癌的发生。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，促进p53降解。我们将野生型p53基因通过腺病毒载体Ad 5CMV-p53导入人宫颈癌细胞系HeLa中，该细胞系含有HPV 18型和野生型p53。细胞在补充有10%热灭活胎牛血清的RPMI培养基中生长。通过将巨细胞病毒启动子、野生型p53 cDNA和SV 40多聚腺苷酸化信号插入到修饰的Ad 5腺病毒的E1缺失区中来产生Ad 5CMV-p53。当使用确保感染复数为100或更大的剂量时，转导效率为100%。免疫组化和Western blot检测到p53蛋白在Ad 5CMV-p53感染的细胞中表达。细胞计数和[3 H]胸苷掺入试验均检测到Ad 5CMV-p53感染细胞的生长受到极大抑制。这些数据表明，转染HPV阳性宫颈癌细胞与野生型p53基因的形式，如Ad 5CMV-p53是一个潜在的新的治疗宫颈癌。
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-typep53into 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-typep53,was used in this study. Cells were grown in RPMI medium supplemented with 10% heat-inactivated fetal bovine serum. Ad5CMV-p53was created by inserting the cytomegalovirus promoter, wild-typep53cDNA, 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-typep53gene in a form such as Ad5CMV-p53is a potential novel therapy for cervical cancer.