Sequence variations and viral genomic state of human papillomavirus type 16 in penile carcinomas from Ugandan patients
Sequence variations and viral genomic state of human papillomavirus type 16 in penile carcinomas from Ugandan patients
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DOI：
10.1099/0022-1317-78-9-2199
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发表时间：
1997-09-01
影响因子：
3.8
通讯作者：
Giraldo, G
中科院分区：
文献类型：
--
作者：
Tornesello, ML;Buonaguro, FM;Giraldo, G
分析了5例乌干达患者阴茎癌活检组织中人乳头瘤病毒16型（HPV-16）E6/E7（nt 34-880）和L-1（nt 6584-7035）ORF以及长对照区（LCR）（nt 7289-93）的序列变异。乌干达是生殖器肿瘤高发国家，根据LCR序列进行系统发育分析，5株分离株均属于African-1谱系（Af 1），而E6基因系统发育分析表明，4株分离株属于Af 1-u亚类，该亚类为Af 1-u亚类。其特征在于位于E6基因5 ′端的三个点突变，导致第10和14位氨基酸的变化，通过在nt 286和289处不存在同义突变而与Afl类区分。5个样品中有3个存在nt 335的非同义替换。E6 Af 1突变模式仅存在于单一的乌干达HPV-16分离株中。E7和L1区的核苷酸序列分析不允许任何Af 1亚类鉴定，这些样品中的病毒DNA的物理状态通过PCR和Southern印迹分析来表征。能够扩增全长E2区的寡核苷酸（nt 2734-3872）未能在五个样品中的四个中扩增靶序列，表明E2 ORF的破坏和HPV基因组整合到人DNA中。Southern印迹分析证实了病毒整合状态，我们的研究结果有助于HPV-16“非洲谱系”的特征与Af iu亚类的鉴定;此外，这也是首次报道显示在男性生殖器癌中HPV-16整合到人类基因组中，并破坏E2 ORF。
Sequence variations in the E6/E7 (nt 34-880) and the L-1 (nt 6584-7035) ORFs, and in the long control region (LCR) (nt 7289-93) of human papillomavirus type 16 (HPV-16) were analysed in five penile carcinoma biopsies obtained from Ugandan patients. Uganda is a country with a high incidence of genital cancers, All five isolates were classified as members of African-1 lineage (Af1) by phylogenetic analysis based on LCR sequences, The E6 gene phylogenetic analysis, however, showed that four isolates fell into a new subclass designated Af1-u, This subclass, characterized by three point mutations located at the 5' end of the E6 gene with resulting changes in amino acids at positions 10 and 14, is distinguishable from the Af1 class by the absence of synonymous mutations at nt 286 and 289. The nonsynonymous substitution at nt 335 was present in three out of five samples. The E6 Af1 mutation pattern was present in only a single Ugandan HPV-16 isolate. Nucleotide sequence analysis of the E7 and L1 regions did not allow any Af1 subclass identification, The physical state of the viral DNA in these samples was characterized by PCR and Southern blot analysis. Oligonucleotides which enable amplification of the full length E2 region (nt 2734-3872) failed to amplify the target sequence in four out of five samples, suggesting disruption of the E2 ORF and integration of the HPV genome into the human DNA, Southern blot analysis confirmed the virus integration status, Our results contribute to the characterization of the HPV-16 'African lineages' with the identification of the Af iu subclass; furthermore, this is also the first report showing that in male genital cancers HPV-16 is integrated into the human genome with disruption of the E2 ORF.