HPV16 E2基因各区域缺失与宫颈病变相关性的研究

目的:检测湖北地区HPV16阳性宫颈标本E2基因碳端(C)、铰链区(H)、氮端(N)的缺失状态,探讨其与CIN及宫颈癌的关系。方法:将122例宫颈标本按病变程度分为宫颈癌组、CINⅡ~Ⅲ组、对照组。应用多重聚合酶链反应(PCR)将各组HPV16感染标本E2基因的C、H、N分别与E6基因同时扩增,检测E2基因各区域的缺失状态,并应用Scion Image4.0软件半定量分析E2、E6基因条带灰度值,判断HPV DNA的整合状态。结果:宫颈癌组E2基因C、H、N缺失率分别是22.5%,50%,77.5%;CIN组为30.77%,69.23%,92.31%;对照组为9.09%,27.27%,27.27%。E2基因C、H缺失率在3组的任两组间比较无统计学差异(P0.05),而E2基因N端缺失率在宫颈癌组与对照组之间及CIN组与对照组之间差异有统计学意义(P均0.01)。结论:HPV16病毒E2基因N端缺失状态与CIN及宫颈癌有内在关系。在检测HPV感染同时检测E2基因N端缺失状态,对间接判断预后及指导后续治疗有重要意义。     

青岛地区不同宫颈病变组织中HPV16 E2基因突变分析

目的:探讨山东省青岛地区不同宫颈病变组织中人乳头瘤病毒16型(HPV16)E2基因序列多态性,以及序列变异与不同宫颈病变的关系。方法:提取257例宫颈脱落细胞及宫颈癌组织DNA,PCR进行HPV分型,其中HPV16阳性标本扩增E2基因,测定PCR产物序列。通过DNAStar生物软件进行核苷酸和氨基酸序列分析。结果:共检测出11个碱基突变位点,其中10个导致氨基酸的改变,突变热点为nt3410(63.2%),nt3159(55.9%),nt3249(55.9%)。统计学分析显示,各个位点的突变与不同级别宫颈病变之间均无相关性(P>0.05)。宫颈癌标本中基因整合率为[65%(14/40)],远高于CINⅢ的整合率[14%(6/43)](P<0.001)。结论:青岛地区HPV16 E2基因存在多个位点的突变,且发现G2828A,T3274G,T3384C,T3524C为青岛地区特异性突变,这些位点的突变可能与高度上皮内瘤变和浸润性宫颈癌的发生和发展有关。     

女性人乳头瘤病毒感染与细菌性阴道病、宫颈病变程度的相关性研究

目的 探讨人乳头瘤病毒（HR-HPV）感染分别与细菌性阴道病、不同宫颈病变程度的相关性。方法 回顾性分析2019年12月至2020年12月在合肥市第二人民医院确诊为细菌性阴道病821例及宫颈病变758例患者的临床资料,按照HR-HPV检测结果分别将其分为阴道HPV阳性组（n=279）与阴道HPV阴性组（n=542）、宫颈HPV阳性组（n=383）及宫颈HPV阴性组（n=375）,分析细菌性阴道病中不同年龄段妇女HPV阳性率及阴道微生态指标;观察不同宫颈病变中HR-HPV的阳性率细菌性阴道病、不同宫颈病变程度之间的相关性;ROC分别评估HR-HPV阳性率在细菌性阴道病与宫颈病变中的诊断价值。结果 细菌性阴道病患者共有279(33.98%)例检查出HR-HPV阳性;阴道微生态检测中HR-HPV阳性患者与阴性患者仅唾液酸苷酶阳性率差异有统计学意义（P <0.05）;宫颈病变患者中共有383例（50.53%）检查出HR-HPV阳性;Spearman显示HR-HPV阳性率与细菌性阴道病、宫颈病变程度之间均呈正相关（P <0.05）;ROC显示HR-HPV阳性在细菌性阴道病及宫颈病变中...     

Brn-3a在宫颈病变中的表达及其与HPV亚型的相关性研究

目的:初步探讨Brn-3a在HPV所致各级别宫颈病变中的表达及不同HPV亚型与Brn-3a表达的关系,探讨其作为CIN诊断和治疗标志物的可行性。方法:按照入组标准选取研究期间在我院就诊的月经正常、有性生活史的妇女97例为研究对象。对同一位患者的宫颈脱落细胞同时进行转录因子Brn-3a的表达及HPV分型的检测。分析每组样本Brn-3a的表达。结果:Brn-3a表达随着病变加重而升高。高级别CIN(CINⅡ、Ⅲ及原位癌)LEEP术后随访组Brn-3a表达高于宫颈良性病变(HPV-)组,差异有统计学意义(P=0.0000,<0.05);该组平均值虽高于HPV感染(HPV+)、宫颈湿疣与CINⅠ组,但二者无统计学差异(P=0.6815,>0.05);与高级别CIN组(治疗前)相比,表达量明显减少,差异有统计学意义(P=0.0002,<0.05)。不同HPV亚型所致的同级别病变中Brn-3a表达差异无统计学意义;高危型和低危型HPV感染所致的CINⅠ以下病变组中,Brn-3a表达无统计学差异(P=0.5815);同种HPV亚型所致的不同病变级别Brn-3a表达有统计学差异。结论:转录因子Brn-3a表达与HPV所致宫颈病变的病理级别相关,并非简单的反映HPV的感染状态。     

妊娠对人乳头瘤病毒16、18感染率及宫颈病变的影响

目的 探讨妊娠对人乳头瘤病毒(HPV)16、18感染率及宫颈病变的影响.方法 2007年1月至11月在哈尔滨医科大学第一临床医学院就诊的已婚育龄妇女30例,分别于妊娠前、妊娠期及产后2个月应用实时荧光定量聚合酶链反应检测人乳头瘤病毒16、18型(FQ-PCR HPV16、18),宫颈脱落细胞薄层液基细胞学检查(TCT),阴道镜检查.结果 观察对象中妊娠晚期的HPV16、18感染率最高为70.0%,妊娠中期次之为33.3%,产褥期下降为20.0%,妊娠早期阳性的2例到妊娠晚期均自然转为阴性.各期HPV感染率比较差异有统计学意义(PO.05).观察过程中无发生流产等不良妊娠结局病例.结论 妊娠增加HPV16、18的感染率,不加重宫颈病变的进展.     

多重人乳头瘤病毒感染检测在宫颈病变诊断中的价值

目的探讨人乳头瘤病毒（human papillomavirus,HPV）检测在宫颈病变筛查中的意义,以及多型别HPV感染与宫颈病变的关系。方法2005年8月至2006年10月应用HPV基因分型技术对北京市垂杨柳医院967例门诊患者进行检测,阳性者进行细胞学检测,两者均阳性者行阴道镜检测;并与同期门诊行细胞学检测的512例患者对比。结果967例中筛查出360例HPV感染患者,感染率为37.23%,单一型别感染率为61.11%,多型别感染率为38.89%。在多型别感染中2种型别感染比例最多（57.86%）。从正常组织到宫颈上皮内瘤样病变（CIN）Ⅱ的不同病变中,多重感染的比例随病变级别上升逐渐增加,与正常者相比差异有显著性意义（P〈0.05）。在CINⅢ和宫颈癌中,多重感染的比例有所下降,但明显高于正常者。HPV联合宫颈薄层液基细胞学（TCT）检测与单纯TCT检测比较,两者中低度鳞状上皮内病变（LSIL）分别为13.33%和5.27%,高度鳞状上皮内病变（HSIL）为4.17%和0.78%,癌为0.56%和0.20%,两者比较差异有显著性意义,P〈0.01。结论结合HPV检测可提高TCT检测的敏感性,多重HPV感染及型别与宫颈病变级别相关。     

HPV及相关基因甲基化在宫颈癌中的研究进展

HPV联合宫颈细胞学筛查后大大提高了宫颈癌前病变的检出率,但它预测宫颈癌发生的价值相对较低。近年,肿瘤相关基因的甲基化已在许多癌症中被确认,对HPV及宫颈癌相关基因甲基化的研究也日益增多。对生物标记物的甲基化分析,使预测宫颈癌的发生和区分病变的严重程度有了新的方法。现将HPV及宫颈癌相关基因甲基化的研究进展作一综述。     

宫颈脱落细胞中的DNA甲基化研究

在宫颈病变发生、发展的过程中,DNA甲基化水平发生一系列改变。以往的研究多是基于宫颈组织的,而近来检测宫颈脱落细胞甲基化的研究越来越多。与宫颈组织相比,用宫颈脱落细胞作为研究对象进行检测安全无创,更方便临床应用,可能成为筛查、诊断宫颈病变和判断宫颈病变预后的有利工具。讨论研究较多的宫颈脱落细胞中基因的甲基化水平与宫颈病变等级的关系及其临床应用。     

人乳头状瘤病毒不同亚型感染与宫颈病变的相关性

目的 了解门诊高危人群和宫颈病变患者的人乳头状瘤病毒（HPV）感染的型别分布，探讨不同HPV亚型感染的致癌性。方法 采用导流杂交技术，对门诊就诊的1715名有性生活史妇女，进行下生殖道HPV感染分型筛查。其中463例经阴道镜下定点官颈活检，病理确诊分为官颈炎症234例、CINⅠ96例、CINⅡ80例和CINⅢ53例。分析人群HPV感染的型别分布、与宫颈病变相关性。结果 21种HPV亚型均被检出，1715名门诊妇女中HPV阳性率57．1％（978／1715），宫颈病变中为76．9％（356／463），而在CINⅢ中达98．1％（52／53）。常见的5种亚型分布：高危人群为HPV16、58、52、33和6型；宫颈病变为HPV16、58、52、33和31型；CINⅢ中为HPV16、33、58、31和52型。HPV16、33和31型与宫颈病变程度相关（P〈0．05），Logistic回归分析，高级别鳞状上皮内瘤变（HSIL，CINⅡ＋CINⅢ）的主要风险也是感染HPV16、33和3l型（OR9．59、2．99和2．52，95％CI4．805～11．989、1．231～7．296和1．174～5．429）。结论 本地区官颈病变的常见型别是HPV16、58、52、33和31型，HPV16、33和31型是HSIL的主要致病型。     

阴道微生态异常与宫颈上皮内病变的相关性研究

目的 探讨阴道微生态异常与宫颈上皮内病变的相关性.方法 选取本院就诊同时行阴道微生态与宫颈活检患者888例,收集年龄、高危型人乳头瘤病毒(HR-HPV)、阴道微生态、孕产史、避孕方式、病理结果等临床资料,分析阴道微生态异常与子宫颈上皮内病变的相关性.结果 888例患者中,232例高级别宫颈鳞状上皮内病变(HSIL)中,...     

Comparison of in vitro translated HPV-16 E7 protein with GST-fusion HPV-16 E7 protein as serologic markers in patients with cervical cancer

Park JS, Um SJ, Kim TY, Kim EJ, Kim CJ, Park SN, Namkoong SE, Kim SJ. Comparison of in vitro translated HPV-16 E7 protein with GST-fusion HPV-16 E7 protein as serologic markers in patients with cervical cancer. Int J Gynecol Cancer 1998; 8 : 380–386.
The purpose of the study is to investigate in vitro translated HPV-16 E7 protein and GST-fusion HPV-16 E7 protein as serologic markers in females with cervical cancer. The sera of 39% (39/100) of patients with cervical cancer were RIPA positive for in vitro translated HPV-16 E7 protein and the sera of 29% (29/100) of females with cervical cancer were ELISA positive for HPV-16 GST-fusion E7 protein. The positivities for in vitro translated HPV-16 E7 protein in cervical cancers were 14% (7/49) in stage I, 59% (26/44) in stage II, and 86% (6/7) in stage III/IV, thus significantly correlating with advancing stage of the disease ( P < 0.01). These data suggest that many patients with cervical neoplasia generate a positive antibody response to HPV-16 E7 proteins. The antibody positives to in vitro translated HPV-16 E7 protein increase with advancing clinical stage of disease. These HPV-16 E7 proteins might be a disease-specific markers which could be useful in adjunctive diagnostic assay of HPV-related cervical neoplasia.     

The prognostic significance of HPV-16 genome status of the lymph nodes,the integration status and p53 genotype in HPV-16 positive cervical cancer: a long term follow up

Objective Prognostic evaluation of HPV-16 genome status of the pelvic lymph nodes, the integration status of HPV-16 and p53 codon 72 polymorphism in cervical cancer.
Design Prospective cohort study.
Setting Department of Gynaecological Oncology, University of Debrecen, Hungary.
Sample Thirty-nine patients with HPV-16 positive cervical cancer.
Methods Primary tumour specimens of 39 cervical cancer patients with HPV-16 positive primary tumour were subjected to multiplex polymerase chain reaction using HPV-16 E1/E2, E7 and p53 codon 72 allele-specific primers. Pelvic lymph nodes of the same patients were also tested for the presence of HPV-16 DNA and for its integration status using HPV-16 E7 and E1/E2 ORF specific primers, respectively.
Main outcome measures Progression-free survival.
Results Metastatic lymph nodes carried HPV-16 DNA more frequently than nodes with no evidence of disease (  100.0% vs 35.7%, P = 0.001  ). Cases with HPV-16 positive nodes had higher recurrence rate than those with HPV-16 negative nodes (  42.9% vs 11.1%, P = 0.009  ). There was no difference between cases with and without histologically proven nodal disease with regard to integration status of HPV-16 DNA in the primary tumour (integrated 90.9% vs 71.4%, episomal 9.1% vs 21.4%, mixed 0% vs 7.1%) and p53 codon 72 polymorphism (Arg/Arg 54.5% vs 67.9%, Pro/Pro 0 vs 7.1%, Arg/Pro 45.5% vs 21.4%).
Conclusion Regardless of the presence of nodal metastasis, HPV-16 status of the nodes is a significant predictor of recurrent disease. HPV-16 integration status and p53 codon 72 genotype do not seem to have a bearing on disease outcome in cervical cancer with HPV-16 positive primary.     

Immune response to p53 and HPV-16 E6 proteins in patients with cervical cancer

Park JS, Kim CJ, Um SJ, Hwang ES, Kim HS, Park SN, Namkoong SE, Kim SJ. Immune response to p53 and HPV-16 E6 proteins in patients with cervical cancer. Int J Gynecol Cancer 1998; 8 : 328–335.
To investigate whether p53 autoantibodies could be found in the sera of patients with cervical cancers, we have therefore studied by radioimmunoprecipitation assay, using in vitro translated p53 protein, sera from such patients. The sero-positive patients for p53 were also evaluated in relation to immunoreactivity to p53 antigens by immunohistochemistry, for genomic alterations of p53 by PCR-SSCP, and for the presence of HPV-16/18 DNAs in the cervical cancer cells. In immunohistochemistry, expression of p53 protein was seen in 47% (14/30) of HPV-16 or -18 positive cervical cancers and 13 % (2/15) of HPV-16/18 negative cervical cancers ( P < 0.01). Eight out of 12 control ovarian cancers (67%) showed positive p53 staining in most tumor cells. Cases of cervical cancer and ovarian cancer, which were positively expressed p53 protein in the tissue or the sera, were studied for genomic alterations in exons 5–8 of the p53 by PCR-SSCP. Serum antibodies to in vitro translated p53 protein were found in two cases from 63 cervical cancers; one patient was stage IIA, having HPV-16 DNA in a tumor of squamous cell type, and another patient was stage IIIB, having HPV-16 and -18 DNAs in an adenocarcinoma. The cervical cancer tissues from the two sero-positive patients were also positive for p53 immunostaining. None of the cervical cancer samples showed aberrant bands, but three of eight cases of ovarian cancer which were positive for p53 protein by immunostaining were shown to have aberrant bands by PCR-SSCP. In contrast to ovarian cancers, alteration of p53 tumor suppressor gene and positive antibody response to p53 protein seem to be rare events in patients with cervical cancer.     

Viral DNA load, physical status and E2/E6 ratio as markers to grade HPV16 positive women for high-grade cervical lesions

OBJECTIVES: Cervical intraepithelial neoplasias (CIN) associated with high-risk (HR) human papillomavirus infection, in addition to HR-HPV typing need other viral marker testing to distinguish a subset of lesions with clinical relevant infections. This study has evaluated the significance of viral markers, such as viral load, physical status and E2/E6 ratio, to stratify HPV16 infected women at a single point in time for grade of cervical lesions. METHODS: One hundred sixty-six cytological specimens were selected from women with low (n=72) and high (n=94) grade squamous intraepithelial lesions (SIL), and positive to HPV16. All the 72 LSIL were CINI, 83 of the 94 HSIL were CINII/III and 11 SCC (Squamous Cervical Carcinoma). Cytological specimens were analysed by two different SYBR Green Real-time PCR assays (RT-PCR). Specific primers for both E2 and E6 viral genes and GAPDH cellular gene were designed to determine viral load, physical status and E2/E6 ratio. RESULTS: The viral load was significantly higher in HSIL than in LSIL. In CINI episomal DNA was prevalent (72.2%), mixed forms (episomal and integrated) were 27.8%, suggestive of an early integration of viral DNA into cellular genome, no pure integrated forms were detected. However in CINII/III mixed DNA forms were prevalent (73.5%). In SCC pure integrated DNA was prevalent (81.8%) in absence of episomal forms. E2/E6 ratio decreased significantly from CINI to CINII/III and SCC with a linear trend. The logistic regression analysis showed that viral load higher than 1.38x10(6) genome copies per 300 ng of total DNA associated with E2/E6 ratio lower than 0.90 was highly significant in differentiating CINII/III versus CINI, while the only E2/E6 value lower than 0.17 was significant in differentiating SCC from CINI. CONCLUSIONS: Viral load higher than 1.38x10(6) genome copies per 300 ng of total DNA and E2/E6 ratio values allow HPV16 infected women with high grade cervical intraepithelial lesions to be recognized.     

Characterization of human papillomavirus genotypes and HPV-16 physical status in cervical neoplasias of women from northern Portugal

Objective

To determine human papillomavirus (HPV) genotypes and the physical status of HPV-16 DNA among women from northern Portugal with cervical lesions.

Methods

The present retrospective study included samples of cervical exfoliated cells from 88 women (median age 42.0 ± 13.1 years) who attended the Gynecology Service at the Portuguese Institute of Oncology in Porto during 2010. After DNA extraction, HPV genotyping was performed by polymerase chain reaction (PCR) followed by restriction fragment length polymorphism analysis using the MY09/MY11 primers. The physical status of HPV-16 was determined by real-time PCR.

Results

Overall, 69.3% of the samples tested positive. The prevalence of HPV infection was 38.5% in normal samples, 57.7% in cervicitis samples, and 87.2% in all cervical lesions including invasive cancers. Sixteen genotypes were detected, the most prevalent ones being HPV-16 (42.9%), HPV-31 (12.2%), and HPV-58 (10.2%); HPV-18 was rare. The overall prevalence of HPV-16 integration was 31.6%. The physical status of HPV-16 did not differ significantly by histology.

Conclusion

The most frequent genotypes were HPV-16, -31, and -58. Integration of HPV-16 DNA seemed to be an early event in cervical carcinogenesis. Further studies are required to clarify the value of viral integration as a prognostic marker.