人乳头瘤病毒与宫颈癌关系研究进展

宫颈癌是一种严重危害女性健康的恶性肿瘤,其发病率较高,位居女性恶性肿瘤的第二位,仅次于乳腺癌。自从1977年德国学者ZurHausen等从宫颈癌标本中发现了人乳头瘤病毒(Human papillomavirus HPV)DNA,并推测HPV感染与宫颈癌发生有关后,许多学者对HPV与宫颈癌的相关性进行了大量的研究,并证实HPV感染是宫颈癌发病的必需因素。目前,对于宫颈HPV感染检测有多种手段,其中聚合酶链反应(PCR)和捕获杂交技术在实验室中应用较广泛。在宫颈癌筛查中联合应用HPV检测和细胞学,不仅可以提高敏感性,而且还可以减少随诊频率,从而大大降低了宫颈癌的发生。     

人乳头瘤状病毒分型检测技术研究进展

HPV病毒有100多个亚型,反复持续的高危型人乳头瘤状病毒(Human papillomavirus,HPV)的感染是子宫颈癌发生的必要条件.对其进行分型检测,对子宫颈病变的早期发现和预防有很重要的意义.近年来,HPV分型检测技术发展十分迅速,本文对其相关研究新进展进行综述.     

高危人乳头瘤病毒基因型与宫颈癌筛查

应用第二代基因杂交捕获技术（HC-Ⅱ）对7068例21-58岁妇女官颈细胞进行13种高危型人乳头瘤病毒（HPV）DNA的检测。该人群中13种高危型HPVDNA的总检出率为23．45％。在检出的1440例HPV阳性者中,宫颈癌、官颈上皮内高度病变（CINⅡ／Ⅲ）及宫颈上皮内低度病变（CINⅠ）的高危型HPVDNA检出率分别为100％、100％、65．06％。女性生殖道高危型HPV感染是宫颈癌及CIN流行的主要危险因素,提示言颈癌的防治应重点预防HPV感染,而对HPV感染的筛查和密切监测应是已感染高危型HPV的对象;     

一氧化氮、人乳头瘤病毒和宫颈癌之间相互关系的研究进展

人乳头瘤病毒感染是宫颈癌发生的重要始动原因,从HPV感染到宫颈癌发生,需要许多共刺激因子的参与。这些共刺激因子均可引起宫颈局部一氧化氮浓度的增高。而一氧化氮既可影响HPV的转录和翻译,又在肿瘤发生过程中具有重要调节作用。深入研究一氧化氮、人乳头瘤病毒感染及宫颈癌之间的关系,可为宫颈癌的防治提供新的重要理论基础和药物研制实验平台,通过使用一氧化氮合酶抑制剂降低宫颈局部NO浓度将为全面有效防治宫颈癌带来新的希望。     

人乳头瘤病毒检测方法研究进展

人乳头瘤病毒（Human papilloma viruses,HPV）是少数几种明确与人类癌症相关的病毒之一,其持续感染是导致宫颈癌发生的重要危险因素。HPV的检测对宫颈癌的筛查及治疗宫颈病变药物的有效性评价有重要作用,因此国内外对HPV的检测方法不断革新。纵观近年来HPV检测方法,主要依赖于分子生物技术检测HPV的DNA、RNA、癌蛋白等,但各种检测方法的原理、适用范围及临床意义均不相同。本文将就目前常用的HPV检测技术及试剂盒进行简要介绍,并对各方法的优缺点、灵敏度及特异性进行分析,以期为临床选择合适的检测方法提供参考。     

金黄色葡萄球菌L型和人乳头瘤病毒与宫颈癌关系的研究

应用改良革兰氏染色和免疫组化染色（ＡＢＣ法）,对２７０例宫颈癌和３３例慢性宫颈炎进行了研究。结果发现,宫颈癌革兰氏染色切片中细菌Ｌ型感染率（７５．１％）明显高于慢性宫颈炎（４５．５％）（Ｐ＜０．０５）,金黄色葡萄球菌Ｌ型抗原阳性率（７６．２％）明显高于人乳头瘤病毒（ＨＰＶ）抗原阳性率（５７．１％）。Ｌ型、ＨＰＶ混合感染率为４２．８％。上述结果表明宫颈癌中金葡菌Ｌ型与ＨＰＶ感染均常见,尤其金葡菌Ｌ型感染更为多见;金葡菌Ｌ型与ＨＰＶ感染的致病特征相似,两者均有不同程度的空泡细胞出现。提示金葡菌Ｌ型感染可能也是宫颈癌的致癌重要因素之一。     

人乳头瘤病毒HPV31型宫颈癌细胞模型的建立

宫颈癌与人乳头瘤病毒感染密切相关,建立宫颈癌实验模型可为宫颈癌的研究提供理想的模拟实验条件。我们通过应用基因重组技术,分别以HPV31型E6和E7基因为目的基因,通过原核表达、蛋白纯化和免疫小鼠等获得其特异性检测抗体。我们还通过构建E6和E7基因真核表达载体、转染C33A细胞、博莱霉素抗性筛选和表达检测等步骤,获得一种稳定的体外宫颈癌细胞系。经酶切鉴定及测序证实细胞基因组已重组插入质粒中的目的基因。我们已成功筛选到稳定的目的mRNA和蛋白表达的阳性细胞系,建立了稳定的人乳头状瘤病毒31型(HPV31)的宫颈癌细胞株,为研究宫颈癌提供了体外实验模型。     

宫颈人乳头瘤病毒感染及其致癌机理的研究进展

人乳头瘤病毒（ＨＰＶ）的感染十分常见,是引起性传播疾病的主要因素之一,同时与宫颈癌有密切关系。本文综述近３～４年来国外文献,以探讨ＨＰＶ感染及其致癌机理。     

宫颈癌患者人乳头瘤病毒(HPV)主要型别及其感染研究

本文探讨了江西省和广东省宫颈癌患者人乳头瘤病毒(Human papillomavirus,HPV)感染及其型别分布,分析了高危型HPV对各种宫颈病变的感染情况,为宫颈癌的早期发现和临床诊治提供科学依据。首先采用细胞学、HPV DNA检测(第二代杂交捕获法,HC2)、电子阴道镜和宫颈化学着色方法筛查宫颈癌患者,经病理镜检确诊,然后用GP PCR-SBT法对宫颈癌患者进行HPV基因分型。江西省溪口镇、古市镇及修水县城宫颈癌癌前病变发生率为5．7‰。HC2方法发现宫颈癌患者13种高危型HPV DNA阳性率为89．9％,宫颈上皮内瘤样病变的为84．8％,对照组为24．5％。采用GP PCR-SBT方法进行基因分型发现,江西省宫颈癌患者存在HPV16、58、31、33、18、66、6、11、56和81十种型别,其中HPV81型在国内外鲜有报道。据此提出生殖道高危型HPV感染是妇女宫颈癌发病的重要因素。并发现江西省宫颈癌高发区妇女高危型HPV感染率为24．5％。建立了HPV基因分型的方法,对HPV致宫颈病变的分子机制进行了分析。     

Identification of Genetic Variants of Human Papillomavirus in a Group of Mexican HIV/AIDS Patients and Their Possible Association with Cervical Cancer

Infections caused by the human immunodeficiency virus (HIV) and human papillomavirus (HPV) cause thousands of deaths worldwide each year. So far, there has been no consensus on whether there is a direct relationship between the incidence of neoplasms and the immunosuppression caused by HIV that could help understand if coinfection increases the likelihood of cervical cancer. The objective of the study was to identify the presence of genetic variants of HPV in a group of HIV-positive women and their possible association with cervical cancer. Cervical samples were taken from HIV-positive patients for cytological analysis to identify the HPV genotype by polymerase chain reaction (PCR) and sequencing. The most prevalent L1 capsid protein mutations in the HPV genotype were analyzed in silico. Various types of HPV were identified, both high-risk (HR) and low-risk (LR). The most prevalent genotype was HPV51. Analysis of the L1 gene sequences of HPV51 isolates showed nucleotide variations. Of the samples analyzed in Puebla, Mexico, HPV51 had the highest incidence (17.5%, 7/40). Different mutations, which could be used as population markers, were detected in this area, and they have not been reported in the L1 databases for HPV51 in Mexico. Genotypes 6, 14, 86, 87, 89, and 91, not detected or reported in samples from patients with HPV in Mexico, were also identified. Data from the population analyzed suggest no direct relationship between HIV immunosuppression and cervical cancer, regardless of the high- or low-risk HPV genotype. Furthermore, it is possible to develop regional population markers for the detection of HPV based on the mutations that occur in the sequence of nucleotides analyzed.     

人乳头瘤病毒(HPV)及其诱发子宫颈癌的新进展

子宫颈癌是一类发病率仅次于乳腺癌并严重危害女性生殖系统健康的最常见的妇科恶性肿瘤,在发展中国家,是女性癌症死亡的第一因素。自德国科学家哈拉尔德.楚尔豪森(Harald zur Hausen)因发现人乳头瘤病毒(Human papillomavirus,HPV)导致子宫颈癌而获得2008年度的诺贝尔生理学或医学奖后,全世界广泛认可人乳头瘤病毒(HPV)是引起子宫颈癌的最主要生物学因素。HPV病毒的持续感染在子宫颈癌的发生、发展过程中起了重要作用。本文就HPV病毒的生物学结构、女性生殖道HPV病毒的感染途径以及HPV病毒诱发子宫颈癌机制的最近的研究进展进行扼要综述。     

Human papillomavirus genotyping by multiplex pyrosequencing in cervical cancer patients from India

Cervical cancer is a leading cause of cancer-related deaths among women in India.Human papillomavirus (HPV) infection is the causative agent of cervical cancer; and infection with the high-risk genotypes, predominantly HPV16 and 18,is the biggest risk factor.Vaccines targeting HPV16 and 18 have been found to confer protection in large- scale clinical trials.HPV genotyping has traditionally been carried out to screen the population "at risk" using indirect methods based on polymerase chain reaction (PCR) using consensus primers combined with various DNA hybridization techniques,and often followed by the sequencing of candidate products.Recently,a high-throughput and direct method based on DNA sequencing has been described for HPV genotyping using multiplex pyrosequencing. We present a pilot study on HPV genotyping of cervical cancer and non-malignant cervical samples using multiplex pyrosequencing.Using genomic DNA from cell lines,cervical biopsies,surgical tissues or formalin-fixed,paraffin- embedded tissue samples,we could successfully resolve 6 different HPV types out of the 7 tested,with their prevalence found to be in agreement with earlier reports. We also resolved coinfections with two different HPV types in several samples. An HPV16 genotype with a specific and recurrent sequence variation was observed in 8 cancer samples and one non-malignant sample. We find this technique eminently suited for high-throughput applications,which can be easily extended to large sample cohorts to determine a robust benchmark for HPV genotypes prevalent in India.     

反向点杂交法快速检测HPV基因型的临床应用

应用反向点杂交法(RDB)的原理,针对HPV 6B, 11, 16, 18, 31, 33和35设计了7条序列作为未标记的特异性寡核苷酸(SSO)探针,分别固定在尼龙膜条上,形成7个点,再与经PCR扩增的样品DNA序列杂交,即可在一个膜条上分辨出这7型HPV中的任一型.此法快速简便,特异性高,不存在假阳性;且因PCR灵敏度高,亦不易出现假阴性.用PCR-RDB法检测保存的宫颈癌组织石蜡包埋标本32例,结果:HPV16阳性22例(68.8%),HPV18阳性5例(15.6%),HPV16/18双重感染2例(6.3%),阴性仅3例(9.3%).     

Therapeutic Efficacy of a VSV-GP-based Human Papilloma Virus Vaccine in a Murine Cancer Model

Human papilloma virus (HPV) infections are associated with almost all cervical cancers and to a lower extend also with anogenital or oropharyngeal cancers. HPV proteins expressed in HPV-associated tumors are attractive antigens for cancer vaccination strategies as self-tolerance, which is associated with most endogenous tumor-associated antigens, does not need to be overcome. In this study, we generated a live attenuated cancer vaccine based on the chimeric vesicular stomatitis virus VSV-GP, which has previously proven to be a potent vaccine vector and oncolytic virus. Genes at an earlier position in the genome more to the 3′ end are expressed stronger compared to genes located further downstream. By inserting an HPV16-derived antigen cassette consisting of E2, E6 and E7 into VSV-GP either at first (HPVp1) or fifth (HPVp5) position in VSV-GP’s genome we aimed to analyze the effect of vaccine antigen position and consequently expression level on viral fitness, immunogenicity, and anti-tumoral efficacy in a syngeneic mouse tumor model. HPVp1 expressed higher amounts of HPV antigens compared to HPVp5 in vitro but had a slightly delayed replication kinetic which overall translated into increased HPV-specific T cell responses upon vaccination of mice. Immunization with both vectors protected mice in prophylactic and in therapeutic TC-1 tumor models with HPVp1 being more effective in the prophylactic setting. Taken together, VSV-GP is a promising candidate as therapeutic HPV vaccine and first position of the vaccine antigen in a VSV-derived vector seems to be superior to fifth position.     

人乳头瘤病毒在宫颈鳞癌宫旁组织中的表达及意义

为研究高危型人乳头瘤病毒(high-risk human papillomavirus,hr-HPV)在宫颈鳞癌宫旁组织中的表达及意义,采用免疫组织化学技术对51例宫颈鳞癌患者术后宫旁组织不同位点的hr-HPV表达情况进行检测,应用重复测量设计的方差分析及单因素分析方法进行统计。hr-HPV在宫旁组织中的表达随着距宫颈距离的增加呈明显的梯度下降趋势(P0.05),在不同临床分期、宫颈浸润深度、淋巴结转移组内表达存在差异(P0.05),当临床分期ⅠB1期、宫颈浸润深度≤1/2以及无淋巴结转移时,hr-HPV在距离宫颈2 cm处主韧带、骶韧带及距离宫颈3 cm处阴道中表达开始与正常对照组无差异(P0.05)。结果表明,部分早期宫颈鳞癌患者可选择Ⅱ型改良式根治性子宫切除术,但阴道切除仍推荐3 cm。     

Some Novel Adenosine Mimics: Synthesis and Anticancer Potential against Cervical Cancer caused by Human Papilloma Virus

Two novel adenosine analogs, viz. 9-(1′-β-D-arabinofuranosyl)-6-nitro-1,3-dideazapurine or Ara-NDDP (1) and 9-(5′-deoxy-5′-S-(propionic acid) (1′-β-D-ribofuranosyl) adenine or SAH analog (2), indigenously synthesized, have been found to be potential anticancer agents against cervical cancer caused by human papilloma virus.