Fletcher K, Macnab J C
Medical Research Council, Institute of Virology, University of Glasgow, Scotland.
Virus Genes. 1989 Aug;2(4):323-33. doi: 10.1007/BF00684040.
We have prepared EMBL3 libraries of DNA extracted from the cervix of a patient with cervical intraepithelial neoplasia (CIN) and isolated seven recombinant clones containing sequences that hybridize to human cytomegalovirus (HCMV) DNA. Restriction analysis of one clone with BamHI and SalI endonucleases revealed that the insert DNA showed a high degree of homology to the HCMV Ad169 genome over the region between the HindIII K/E site and the SalI site located within the BamHI P fragment. The HCMV insert in the CIN clone is integrated and flanked by cellular sequences. The major immediate early gene that encodes a polypeptide of approximately 69 kD was found to be conserved in the CIN clone. Transfection of clones encoding the immediate early region of HCMV resulted in cells that were positive in immunofluorescence studies with two monoclonal antibodies directed against the HCMV 69 kD immediate early polypeptide. Infection of human ectocervical cells with HCMV Ad169 revealed that they could express the 69 kD polypeptide encoded by the immediate early gene but could not replicate the virus, whereas HCMV was able to replicate productively in cultured endocervical cells. HCMV has been shown to activate endogenous retroviruses and also to transcriptionally activate the long terminal repeat of human immunodeficiency virus. Activation of virus and cellular genes by HCMV may be a means by which this virus is involved in the multistage process of oncogenesis and/or the activation of latent infections.
我们制备了从一名宫颈上皮内瘤变（CIN）患者宫颈中提取的DNA的EMBL3文库，并分离出7个重组克隆，这些克隆包含与人类巨细胞病毒（HCMV）DNA杂交的序列。用BamHI和SalI内切酶对其中一个克隆进行限制性分析表明，插入DNA在位于BamHI P片段内的HindIII K/E位点和SalI位点之间的区域与HCMV Ad169基因组具有高度同源性。CIN克隆中的HCMV插入片段整合在细胞序列之间。发现编码约69kD多肽的主要立即早期基因在CIN克隆中是保守的。用针对HCMV 69kD立即早期多肽的两种单克隆抗体进行免疫荧光研究，结果显示转染编码HCMV立即早期区域的克隆后细胞呈阳性。用人宫颈外细胞感染HCMV Ad169表明，它们可以表达由立即早期基因编码的69kD多肽，但不能复制病毒，而HCMV能够在培养的宫颈内细胞中高效复制。已证明HCMV可激活内源性逆转录病毒，还可转录激活人类免疫缺陷病毒的长末端重复序列。HCMV对病毒和细胞基因的激活可能是该病毒参与肿瘤发生多阶段过程和/或潜伏感染激活的一种方式。
