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Molecular variants of human papillomavirus type 16 and 18 and risk for cervical neoplasia in Portugal 总被引:1,自引:0,他引:1
Pista A Oliveira A Barateiro A Costa H Verdasca N Paixão MT 《Journal of medical virology》2007,79(12):1889-1897
Persistent high-risk human papillomavirus (HPV) infection is considered as the central cause of invasive cervical cancer. Specific HPV 16 and 18 sequence variations were associated with an increased risk for progression. The purpose of this study was to analyze intratypic variations of HPV 16 and 18 within the E6 gene, MY09/11 and LCR regions, and to evaluate the risk of these variants for cervical neoplasia among Portuguese women. Cervical samples from 187 HPV 16-positive and 41 HPV 18-positive women with normal epithelium, cervical intraepithelial neoplasia, or invasive cervical cancer were amplified by type-specific PCR, followed by sequence and phylogenetic analysis. Sixteen new HPV 16 and 18 patterns are described in this paper. European HPV 16 variants were the most frequent (74.3%), particularly Ep-T350 (44.4%), followed by African (16.1%), and Asian-American (9.6%). Non-European HPV 16 variants were more frequent in pre-invasive lesions than in normal tissue and low-grade lesions. However, when analyzed separately, only African variants were associated significantly with an increased risk for cervical cancer. For HPV 18, the AsAi variant showed a trend, which was not statistically significant to an enhanced oncogenicity. European variants seemed to be significantly associated with a lower risk for cervical cancer development. The distribution of HPV 16 and 18 variants was not related to age or race among women living in the same geographical region. Knowledge of variants will be important for risk determination as well as for designing primers or probes for HPV detection methods, and for appropriate cervical cancer prevention strategies. 相似文献
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目的检测HPV16/18和HPV16E6/E7 DNA在宫颈癌组织中的表达,探讨其在宫颈癌发病中的作用.方法应用PCR和琼脂糖凝胶电泳方法检测46例宫颈癌组织中HPV16/18和HPV16E6/E7DNA.结果 46例宫颈癌中56.5%(26/46)扩增HPV16/18 DNA,其中宫颈鳞癌25例,宫颈腺癌1例.正常对照组20例HPV16/18DNA均为阴性,与宫颈癌组相比差异有显著性(P<0.01).HPV16/18 DNA阳性拷贝对数值为4.32±2.45.HPV16E6,E7DNA分别有53.8%(14/26)、46.2%(12/26)扩增.结论 HPV16/18和HPV16E6/E7 DNA与宫颈癌的发生密切相关,是宫颈癌恶性转化的关键之一,预示着宫颈癌有较强的增殖能力和转移能力. 相似文献
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Michiko Miyashita Dorothy M. Agdamag Toshiyuki Sasagawa Kaori Matsushita Lourdes Ma. Salud Calixto O. Salud Kunikazu Saikawa Prisca S. Leano Teresita Pagcaliwagan Jessica Acuna Azumi Ishizaki Seiji Kageyama Hiroshi Ichimura 《Journal of medical virology》2009,81(3):545-551
In order to prevent cervical cancer, vaccines against human papilloma virus types 16 (HPV‐16) and 18 (HPV‐18) have been implemented worldwide. However, the HPV types that cause cancer can differ according to geographical area and ethnicity. In this new era of the HPV vaccine, it is important to elucidate the prevalent HPV types in each area. Therefore, the prevalence of HPV infection and cervical abnormalities among 369 female commercial sex workers in the Philippines were examined. HPV L1 gene was amplified by polymerase chain reaction (PCR) using modified GP5+/6+ primers, and genotyping was performed by sequencing cloned PCR products. HPV DNA was detected in 211 (57.2%) women, among whom 46 HPV types were identified. HPV‐52 was most common and multiple‐type infection was observed in 44.5%. Among 56 women with abnormal cervical cytology (low‐ and high‐grade squamous intraepithelial lesions and adenocarcinoma in situ), HPV‐52 was most common (23.2%), followed by HPV‐16 (19.6%), ‐58 (10.7%), and ‐67 (10.7%). Only 27% of these women were positive for HPV‐16 and ‐18. Multivariate analysis revealed that HPV‐16, ‐39, ‐52, ‐67, and ‐82 were significantly associated with abnormal cytology. Repeated analysis of HPV‐52 single‐positive samples using the original GP5+/6+ PCR primers produced negative results in 57% of cases, suggesting that the prevalence of HPV‐52 infection may have been underestimated in previous studies, and the current vaccines may not be sufficient for preventing infection and the development of premalignant lesions of the cervix in women in the Philippines. J. Med. Virol. 81:545–551, 2009. © 2009 Wiley‐Liss, Inc. 相似文献
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目的 用基因重组技术构建pcDNA-E6E7真核表达载体.方法 经限制性内切酶和序列分析,用脂质体转染技术将其转入B16细胞,G418稳定筛选后IFA法检测其表达,RT-PCK法检测HPV16E6E7mRNA的生成,并将转染细胞接种小鼠皮下,观察成瘤情况.结果 酶切鉴定证实重组质粒中插入的目的基因片段及载体大小、方向和插入住点均正确,在转染的B16细胞中可见绿色荧光并检测到HPV16E6E7mRNA的生成,接种的转染细胞在小鼠皮下100%成瘤.结论 提示B16细胞转染E6E7后其致瘤性与转染空载体组和野生型B16细胞组无明显差异. 相似文献
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HPV16 E6 natural variants exhibit different activities in functional assays relevant to the carcinogenic potential of E6 总被引:5,自引:0,他引:5
Lichtig H Algrisi M Botzer LE Abadi T Verbitzky Y Jackman A Tommasino M Zehbe I Sherman L 《Virology》2006,350(1):216-227
Genetic studies have revealed natural amino acid variations within the human papillomavirus (HPV) type 16 E6 oncoprotein. To address the functional significance of E6 polymorphisms, 10 HPV16 E6 variants isolated from cervical lesions of Swedish women were evaluated for their activities in different in vitro and in vivo assays relevant to the carcinogenic potential of E6. Small differences between E6 prototype and variants, and among variants, were observed in transient expression assays that assessed p53 degradation, Bax degradation, and inhibition of p53 transactivation. More variable levels of activities were exhibited by the E6 proteins in assays that evaluated binding to the E6-binding protein (E6BP) or the human discs large protein (hDlg). The E6 prototype expressed moderate to high activity in the above assays. The L83V polymorphism, previously associated with risk for cancer progression in some populations, expressed similar levels of activity as that of the E6 prototype in most functional assays. On the other hand, L83V displayed more efficient degradation of Bax and binding to E6BP, but lower binding to hDlg. Results of this study indicate that naturally occurring amino acid variations in HPV16 E6 can alter activities of the protein important for its carcinogenic potential. 相似文献
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The Arg/Arg genotype versus Arg/Pro or Pro/Pro at codon 72 of the p53 gene in association with human papillomavirus (HPV) 16 E6 variants has been implicated as a risk marker in cervical neoplasia. However, research on this topic has produced controversial results. The association of p53 codon 72 polymorphism alone and in combination with specific HPV 16 E6 variants with risk of developing squamous intraepithelial cervical lesion has been investigated in low and high‐grade squamous intraepithelial lesions and in HPV‐negative controls from an Italian population. The data obtained showed statistically significant different distribution of p53 genotypes between healthy controls and precursor lesions, with the p53 arginine homozygous increased in high‐grade squamous intraepithelial lesions. The T350G HPV 16 variant was the most frequent variant observed in the analyzed group of Italian women, showing a slight decreasing with the severity of the lesion. At the same time, the number of the prototype T350 slightly increased with the severity of the cytological lesions. In conclusion, p53 arginine homozygous was found to be increased in high‐grade lesions, supporting the results of previous investigations indicating that HPV‐positive patients with p53 Arg/Arg have an increased risk of developing pre‐cancerous lesions. In addition, T350G HPV 16 variant was over‐represented in p53 Arg homozygous women with cervical lesions. When p53 genotype and HPV 16 variants are considered together, no difference emerges between cases and controls so is not possible to assess that the oncogenic effect of HPV 16 T350G variant may be influenced by the p53 genotype. J. Med. Virol. 85:83–90, 2012. © 2012 Wiley Periodicals, Inc. 相似文献
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Asadurian Y Kurilin H Lichtig H Jackman A Gonen P Tommasino M Zehbe I Sherman L 《Journal of medical virology》2007,79(11):1751-1760
Genetic variations in the E6 oncogene have been associated with different risk for cancer progression. In the present study, the functional significance of human papillomavirus (HPV) polymorphism in the E6 oncogene was investigated. Ten HPV16 E6 variants containing amino acid substitutions in the N-terminal region of E6 were evaluated for different biological and biochemical activities in human keratinocytes, the target cells for HPV infection. Western blot analyses of primary foreskin human keratinocytes or immortalized human keratinocytes, stably transduced with the E6 variants, revealed reduced p53 and Bax levels in all E6 expressing cultures. The reduction induced by most E6 proteins was at similar levels and comparable to the reduction induced by the E6 prototype. The ability of the proteins to induce serum/calcium-differentiation resistant colonies in primary keratinocytes was more variable. Overall activities of the variants ranged between 0.24- and 2.18-fold of the E6 prototype activity. The I27R/L83V variant showed the lowest activity whereas the R8Q variant showed the highest activity. The L83V polymorphism previously associated with risk for cancer progression in some populations, showed significant activity, comparable to that of the E6 prototype, in reducing p53 and Bax levels. Furthermore, this variant showed enhancement in the ability to induce colonies resistant to serum/calcium-triggered differentiation, however, the difference from the prototype was not statistically significant. This, and augmentation of other described functions might result in differences in L83V pathogenicity. 相似文献
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Valeria Cento Massimo Ciccozzi Luigi Ronga Carlo Federico Perno Marco Ciotti 《Journal of medical virology》2009,81(9):1627-1634
High‐risk human papillomaviruses (HPVs) are risk factors for the development of cervical cancer. HPV 16 is the most common type, being present in about 60% of cervical cancers worldwide. Previous studies have reported upon the association between HPV 16 E6 variants and increased risk of cervical intraepithelial neoplasia and invasive cervical cancer. In this study, the presence of HPV 16 polymorphisms in the E6, E7, and L1 genes was investigated in relation to the presence of high‐grade lesions. Sequencing of the E6 gene revealed the presence of nucleotide mutations resulting in 15 amino acid changes. Of these, the G134D and C136R fall within the CXXC zinger finger domain important for p53 binding. In the E7 gene, four nucleotide variations were identified with two leading to the amino acid substitutions L15V and S31R. The L1 gene showed 13 nucleotide changes leading to 11 amino acid substitutions. Among these, the R364C and N367D are located at the base of the HI‐loop of the L1 protein, considered to be the immunodominant epitope of HPV 16. No significant relationship between HPV 16 variants and high‐grade lesions was found. Phylogenetic analysis showed that all the HPV 16 variants identified belonged to the European lineage, except one which was of the Asian‐American lineage. The European‐350G variant was detected most frequently (22 of 34, 64.7%). The study provides some new data on the genetic diversity of HPV 16 which may help to understand the oncogenic potential of the virus and to improve management of patients. J. Med. Virol. 81:1627–1634, 2009. © 2009 Wiley‐Liss, Inc. 相似文献
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中国地方株人乳头瘤病毒16型E7基因一级结构及其变异 总被引:6,自引:0,他引:6
从湖北地区一宫颈癌活检组织中提取DNA,采取加端聚合链反应(Add-on PCR)技术,获得了人乳头瘤病毒16型E7基因(HPVI6E7)。将该基因克隆于载体pUC18后,进行了该基因一级结构顺序分析。完整的HPV16E7湖北株基因(HPV16E7-HB)全长294bp,与已发表的德国株(GS)大小一致,但其核苷酸顺序中有2处发生了变异,均为C→T变异。第43位CAA→TAA使相应的谷氨酰胺密码子变为终止密码,形成无义突变(Nonsensemutation)。将重组质粒中0.3kb的HPV16E7基因在表达载体PWR590-1中进行克隆,经诱导使重组表达质粒在大肠杆菌中高效表达,得到了预计的、分子量约为69×10~3的融合蛋白。该蛋白的表达量占菌体总蛋白量的30%左右。该试验表明HPV16E7一级结构以及所编码的蛋白多肽在不同的国家和/或不同的地区可能存在着差异。本文首次报道了中国地方株HPV16E7基因的一级结构。 相似文献
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KrennHrubec K Mrad K Sriha B Ben Ayed F Bottalico DM Ostolaza J Smith B Tchaikovska T Soliman AS Burk RD 《Journal of medical virology》2011,83(4):651-657
Cervical cancer is the second most common cancer among Tunisian women, and the incidence rates vary by region. Three Tunisian registries report age-standardized rates of 6.3/10(5) in the central region, 5.4/10(5) in the north, and 2.7/10(5) in the south. High-risk human papillomavirus (HPV) types and their variants differ in carcinogenic potential and geographic distribution. The HPV type and variant distribution could be a factor in the differing rates between regions of Tunisia. Tumor tissue was collected from 142 Tunisian cervical cancer patients. Demographic and reproductive characteristics of the patients were abstracted from cancer registry and hospital records. HPV type and variant analyses were performed using PCR-based Luminex and dot-blot hybridization assays. Eighty-three percent of tumors were infected with at least one HPV type. European variants of HPV16/18 were the most prevalent in tumors from all three regions, with all HPV18 infections and 64% of HPV16 infections being of European lineage. A higher frequency of HPV16 was present in Northern Tunisia (80%) than in Central (68%) or Southern Tunisia (50%) (P=0.02). HPV18/45 was significantly more common in adenocarcinomas (50%) than in squamous cell carcinomas (11%) (P=0.004). Frequent infection with European HPV variants most likely reflects the history of European migration to Tunisia. In addition to the importance of understanding the variants of HPV in Tunisia, behavioral and cultural attitudes towards screening and age-specific infection rates should be investigated to aid the development of future vaccination and HPV screening programs and policies. 相似文献
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Human T-cell lymphotropic virus in a population of pregnant women and commercial sex workers in South Western Nigeria
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Background: Over 20 million persons are infected with HTLV-I/II globally. The virus is endemic in Africa and it is also transmitted sexually. Continued identification of high risk groups is important for the control of the disease. Objectives: To determine the prevalence of HTLV infection amongst two highly sexually active groups, pregnant women and CSWs in South Western Nigeria. Methods: Serum samples were tested for the presence of HTLV-I/II antibodies using the Vironostika((R)) HTLV-I/II micro ELISA system. Results: A total of 364 serum samples collected from pregnant women, commercial sex worker (CSW) and secondary school students (control group) from Ibadan. While only 4 (5.1%) of 78 secondary school students (average age: 13years) were reactive for HTLV infection, 20 (16.7%) of 120 pregnant women (average age: 26years) and 38 (22.9%) of 166 CSWs (average age: 23years) were found to have antibodies against HTLV in their sera. The results of this study thus show that HTLV infection is active in the population although higher in pregnant women (although not statistically significant) and CSWs (p>0.05). Pregnant women and CSWs are therefore at a higher risk of HTLV transmission than other members of the population. Conclusion: Routine screening for HTLV infection may go a long way to understanding the epidemiology of HTLV infection in Nigeria and subsequently provide tools for its prevention and control. 相似文献
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新疆妇女子宫颈病变组织中HPV16E6基因突变分析 总被引:2,自引:2,他引:2
目的通过检测HPV16E6基因突变在新疆维吾尔族和汉族妇女宫颈癌组织中的分布规律,以探讨该突变与宫颈癌发生的关系。方法取汉族宫颈炎(含石蜡包埋组织及宫颈刷液基标本)125例,维吾尔族宫颈炎(含石蜡包埋组织及宫颈刷液基标本)124例;汉族石蜡包埋宫颈癌35例,维吾尔族石蜡包埋宫颈癌共109例。用以上HPV16阳性DNA模板PCR扩增HPV16E6全长基因,PCR产物直接测序,分析新疆女宫颈癌组织HPV16E6基因的突变。结果PCR检测结果显示汉族族宫颈炎组织中HPV16E6阳性率为35.71%(15/42);维吾尔族宫颈炎组织中HPV16E6阳性率为30.46%(14/46);汉族族宫颈癌组织中HPV16E6阳性率为33.33%(2/6),维吾尔族宫颈癌组织中HPV16E6阳性率为22.22%(12/54);宫颈炎与宫颈癌中HPV16E6阳性表达无统计学意义(P>0.05),对21份(上皮内低度病变1例,原位癌1例,低分化宫颈癌4例,中分化宫颈癌4例,高分化1例,轻度炎症4例,中度炎症3例,重度炎症1例,正常宫颈2例)HPV16E6扩增片段的双向测序,其中5例成功测序得到一级结构,并且序列分析表明,2例(维吾尔... 相似文献
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Sonia Perez Castro Amparo Iñarrea Fernández María José Lamas González María Teresa Sarán Diez Ana Cid Lama María Jesús Alvarez Martín Mónica Pato Mosquera Isabel López‐Miragaya Nuria Estévez Julio Torres Piñón María Oña Navarro 《Journal of medical virology》2013,85(6):1063-1068
High‐risk human papillomavirus (HPV) DNA detection provides high sensitivity but low specificity for moderate‐grade cervical intraepithelial neoplasia or worse histological identification. A prospective study evaluated mRNA testing efficacy for predicting this histological diagnosis in case of HPV 16 and/or 18 DNA detection. A total of 165 endocervical samples harboring HPV 16 and/or 18 DNA were tested with NucliSENS‐EasyQ® HPV E6/E7‐mRNA‐assay (Biomerieux, Marcy l´Etoile, France). Women with cytological alterations were referred to colposcopy (n = 111). Moderate‐grade cervical intraepithelial neoplasia or worse was diagnosed in 25.8% of women presenting atypical squamous cells of undetermined significance or low‐grade squamous intraepithelial lesions and in 89.8% of women with high‐grade squamous intraepithelial lesions. mRNA sensitivity was 81.3% and 84.1%, respectively. Specificity was 52.2%, and 80.0%, respectively. Negative predictive value (NPV) was 88.9% in undetermined or low‐grade squamous lesions. Positive predictive value (PPV) was 97.4% in high‐grade squamous lesions. mRNA reduced colposcopies by 44.3% in undetermined or low‐grade squamous lesions. Direct treatment of mRNA‐positive cases reduced 77.5% of colposcopies in high‐grade squamous lesions. Women without cytological alterations were followed for 18 months (n = 35), and moderate‐grade cervical intraepithelial neoplasia or worse was diagnosed in 34.3%; mRNA sensitivity and specificity were 83.3% and 86.9%, respectively. PPV and NPV were 76.9% and 90.9%, respectively for predicting moderate‐grade cervical intraepithelial neoplasia or worse in 18 months. mRNA reduced the number of visits for follow‐up in 62.2%. In conclusion, NucliSENS‐EasyQ® HPV E6/E7‐mRNA‐assay (Biomerieux) can serve as a triage test in case of HPV 16 and/or 18 DNA detection. J. Med. Virol. 85: 1063–1068, 2013. © 2013 Wiley Periodicals, Inc. 相似文献
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Association between human papillomavirus type 16 E6 and E7 variants with subsequent persistent infection and recurrence of cervical high‐grade squamous intraepithelial lesion after conization
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Lei Zhang Binlie Yang Ai Zhang Aizhi Zhou Jieyan Yuan Yuhua Wang Liyan Sun Huimin Cao Jieru Wang Wenxin Zheng 《Journal of medical virology》2016,88(11):1982-1988
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pcDNA3/HPV16 E6真核表达质粒的构建及裸DNA注射动物实验观察 总被引:1,自引:0,他引:1
目的探讨HPV16E6基因DNA诱发体液和细胞免疫反应的能力。方法利用基因工程技术构建了HPV16E6真核表达质粒pcDNA3/E6,脂质体法转染Cos7细胞,肌肉注射免疫BALB/c小鼠,免疫组化技术检测抗体产生及抗原表达。结果被转染的Cos7细胞表达HPV16E6蛋白免疫小鼠产生抗HPV16E6抗体。结论这一结果为HPV16相关宫颈癌治疗性DNA疫苗的研制提供了资料。 相似文献