Pater M M, Pater A
Basic Medical Sciences, Faculty of Medicine, Memorial University of Newfoundland, St. John's, Canada.
J Med Virol. 1988 Oct;26(2):185-95. doi: 10.1002/jmv.1890260210.
A total of six cervical carcinoma cell lines, of which two contained human papillomavirus (HPV) type 16 and four contained HPV type 18 DNA, were examined for viral RNA expression by spot-blot hybridization. All six cell lines contained HPV-specific cytoplasmic poly(A)-containing RNA. The same RNA species were detected in both cytoplasmic and polysomal fractions, indicating nonselective translation of these messages into protein products. Primer extension analysis of RNA from the cell lines revealed the presence of three major start sites at the 5' end of the E6 gene for RNA from HPV 16-containing cell lines and four major start sites at the 5' end of the E6 gene for RNA from HPV 18-containing cell lines. One cell line, ME 180, had a distinct pattern with only one minor HPV 18 start site. Northern-blot hybridization using the E6-E7 region of HPV 16 and 18 DNAs as probes revealed the presence of E6-E7 transcripts in all the cell lines except ME 180. Southern-blot analysis revealed that while sequences hybridizing to the E6-E7 coding region were present, the 5' end region containing the enhancer-promoter region of viral DNA is deleted in ME 180. Subsequent Northern-blot hybridization using the E1 region of HPV 18 DNA as probe revealed that the viral RNA in ME 180 is transcribed from this region.
通过斑点杂交对总共六种宫颈癌细胞系进行了病毒RNA表达检测，其中两种含有16型人乳头瘤病毒（HPV），四种含有18型HPV DNA。所有六种细胞系均含有HPV特异性的细胞质含多聚腺苷酸（poly(A)）RNA。在细胞质和多核糖体组分中均检测到相同的RNA种类，这表明这些信息可无选择性地翻译为蛋白质产物。对这些细胞系的RNA进行引物延伸分析发现，含16型HPV的细胞系中，E6基因5'端有三个主要起始位点；含18型HPV的细胞系中，E6基因5'端有四个主要起始位点。有一种细胞系ME 180呈现出不同的模式，只有一个次要的18型HPV起始位点。以16型和18型HPV DNA的E6 - E7区域为探针进行Northern杂交分析发现，除ME 180外，所有细胞系中均存在E6 - E7转录本。Southern杂交分析表明，虽然存在与E6 - E7编码区域杂交的序列，但ME 180中病毒DNA包含增强子 - 启动子区域的5'端区域缺失。随后以18型HPV DNA的E1区域为探针进行Northern杂交分析发现，ME 180中的病毒RNA是从该区域转录而来的。
