宫颈病变中双链RNA依赖的蛋白激酶与HPV感染的相关性

目的：通过检测双链RNA依赖的蛋白激酶（PKR）、人乳头瘤病毒（HPV）在宫颈病变组织中的表达,探讨PKR在宫颈病变发生发展中的意义及可能机制。方法：选择临床病历资料和病理资料完整的宫颈癌患者共37例、同期宫颈上皮内瘤样病变（CIN）Ⅰ～Ⅲ患者56例为研究对象,因子宫肌瘤等良性病变切除子宫的正常宫颈组织30例为对照。采用免疫组织化学SABC法检测PKR及HPV16/18 E6的表达,分析二者与宫颈病变发生发展的关系。结果：宫颈癌组织中PKR表达阳性率为54.1%,高于CINⅢ,CINⅠ～Ⅱ组和正常宫颈组,差异有统计学意义（P <0.05）;在宫颈癌组中PKR表达与HPV16/18 E6表达存在负相关 （r =-0.554,P <0.05）;在CINⅢ组中PKR表达与HPV16/18 E6表达存在正相关（r =0.480,P =0.032）。结论：PKR的异常表达在CINⅢ向宫颈癌发展过程中可能发挥主要作用,此外SABC法测定PKR和E6的表达有可能成为筛选子宫颈癌的早期指标。     

子宫颈癌组织中人乳头状瘤病毒16 E6 mRNA表达与survivin蛋白表达的相关性

目的探讨宫颈癌组织中人乳头状瘤病毒(HPV)16E6mRNA表达与survivin蛋白表达的相关性。方法采用半定量PCR技术和免疫组化链霉菌抗生物素蛋白-过氧化物酶连接法,检测慢性宫颈炎、宫颈上皮内瘤变(CIN)及宫颈癌共148例患者宫颈组织中HPV16E6mRNA及survivin蛋白的表达。结果148例患者中,HPV16阳性共37例,其中慢性宫颈炎5例、CINⅠ6例、CINⅡ～Ⅲ11例及宫颈癌15例。慢性宫颈炎、CINⅠ、CINⅡ～Ⅲ及宫颈癌组织中,HPV16E6mRNA的表达水平分别为0.3±0.4、0.6±0.4、1.8±0.6及2.4±0.6;survivin蛋白阳性表达率分别为7%、31%、63%及84%。CINⅡ～Ⅲ及宫颈癌组织中,HPV16E6mRNA的表达水平及survivin蛋白阳性表达率均显著高于慢性宫颈炎及CINⅠ组织,两者比较,差异均有统计学意义(P<0.01)。宫颈癌组织中,HPV16E6mRNA的表达水平与survivin蛋白阳性表达率呈显著正相关关系(γs=0.62,P<0.05)。结论HPV16E6mRNA的表达水平与宫颈病变的进展有关,survivin蛋白阳性表达率升高可能与宫颈癌的发生、发展密切相关。     

HPV16/18基因DNA存在状态与宫颈病变发展的研究

目的:研究人乳头瘤病毒(HPV)16/18两种亚型的DNA在宿主细胞中的存在状态,探讨其与宫颈病变发生发展之间的关系。方法:2016年1月至2018年2月于北京妇产医院行宫颈脱落细胞检查结果为意义不明的不典型鳞状细胞(ASC-US)及以上病变并经快速导流杂交法明确为HPV16/18型感染的宫颈组织共156例。实时定量PCR扩增检测HPV16/18早期基因E2、E6/E7的拷贝数,分析病毒的整合状态。结果:随着宫颈病变级别的增高,HPV16 E2基因的不完整率逐渐增加,但各组比较无统计学差异(OR=0.625,95%CI=0.151~2.586,P>0.05,r=0.113)。HPV18阳性炎症组织中HPV常以游离形式存在(66.7%),全部宫颈癌组织中HPV18均以完全整合状态存在。随着HPV16/18感染的宫颈脱落细胞判读级别的升高,HPV DNA整合率升高(P<0.05)。结论:E2基因的断裂在HPV16/18感染后的早期事件中很常见。HPV16基因整合可能是其持续感染的主要原因,也是其感染导致宫颈癌的一个重要危险因素。HPV18相关病变常很快的经癌前病变的阶段而直接发展为宫颈癌,推断HPV-18感染更具侵袭性。     

宫颈癌及其癌前病变组织中FHIT蛋白异常表达与HPV16E6、HPV16E7蛋白表达的相关性

目的 探讨宫颈癌中三联脆组(FHIT)蛋白表达与HPV16 E6、E7蛋白表达的相关性.方法 采用免疫组化SP法对四川大学华西第二医院1999年1月至2003年2月的15例正常宫颈、25例宫颈上皮内瘤变(CIN)以及61例浸润性宫颈鳞癌组织标本进行FHIT蛋白、HPV16E6、HPV16 E7蛋白表达的检测.结果 (1)在正常宫颈上皮、CINI～II、CINⅢ及浸润性宫颈鳞癌中,FHIT 蛋白阳性表达率分别为100%(15/15)、71.43%(10/14)、36.36%(4/11)、14.75%(9/61),P＜0.05;HPV16E6蛋白阳性表达率分剐为0(0/15)、7.14%(1/14)、36.36%(4/11)、59.02%(36/61),P＜0.05;HPV16E7蛋白阳性表达率分别为20.00%(3/15)、42.86%(6/14)、63.64%(7/11)、57.38%(35/61),P＞0.05.(2) 宫颈病变组织中FHIT蛋白的阳性表达与HPV16E6蛋白阳性表达呈负相关(P＜0.0l,r=-0.449),与HPV16E7蛋白表达无相关性(P＞0.05).结论 宫颈癌中FHIT 蛋白的异常表达与HPV16 E6蛋白表达有关,FHIT蛋白和HPV16E6蛋白的联合检测可能可作为宫颈癌前病变转归的指标.     

ET-1、ETA-R在宫颈癌中的表达与HPV感染的关系

目的:探讨ET-1及ETA-R表达在宫颈癌发生中的病理生理作用及其与HPV感染的关系。方法:应用免疫组化SP法对68例宫颈癌组织及10例非癌变宫颈组织进行ET-1、ETA-R、HPV16/18E6免疫特异性染色。结果:(1)HPV16/18E6阳性率在宫颈癌中占82.3%,对照组宫颈组织中占20%,二组HPV16/18E6表达有统计学差异(P<0.001);(2)HPV阳性宫颈癌组织ET-1、ETA-R阳性表达率100%,而HPV阴性宫颈癌组织ET-1、ETA-R阳性表达率0%,对照组ET-1、ETA-R阳性表达率20%,3组ET-1、ETA-R表达有统计学差异(P<0.001);(3)ET-1、ETA-R在宫颈癌中的表达均与临床分期、病理分级、组织学类型无关(P>0.05)。结论:HPV是否感染与功能性ET-1/ETA-R自分泌环有关,而ET-1/ETA-R自分泌环是肿瘤生长的驱动器,其功能上调可能是HPV感染子宫颈上皮细胞引起宫颈癌发生的一种新的调控机制。     

宫颈病变中双链RNA依赖的蛋白激酶与HPV感染的相关性

目的：通过检测双链RNA依赖的蛋白激酶（PKR）、人乳头瘤病毒（HPV）在宫颈病变组织中的表达,探讨PICR在宫颈病变发生发展中的意义及可能机制。方法：选择临床病历资料和病理资料完整的宫颈癌患者共37例、同期宫颈上皮内瘤样病变（CIN）Ⅰ-Ⅲ患者56例为研究对象,因子宫肌瘤等良性病变切除子宫的正常宫颈组织30例为对照。采用免疫组织化学SABC法检测PICR及HPV16/18E6的表达。分析二者与宫颈病变发生发展的关系。结果：宫颈癌组织中PKR表达阳性率为54．1％,高于CINⅢ。CINⅠ-Ⅱ组和正常宫颈组,差异有统计学意义（P〈O．05）;在宫颈癌组中PICR表达与HPV16／18E6表达存在负相关（r=-0．554,P〈0．05）;在CINⅢ组中PKR表达与HPV16／18E6表达存在正相关（r=0．480,P=0．032）。结论：PKR的异常表达在CINⅢ向宫颈癌发展过程中可能发挥主要作用．此外SABC法测定PKR和E6的表达有可能成为筛选子宫颈癌的早期指标。     

宫颈癌及癌前病变组织中Notch1及HPV16 E6/E7表达的研究

目的 探讨Notch1受体和人乳头瘤病毒16感染在宫颈癌前病变和宫颈癌发生发展中的作用。方法 采用免疫组化SP法检测18例宫颈上皮内瘤变（cervical intraepithelial neoplasia,CIN）和35例宫颈癌标本中Notch1受体及HPV16E6/E7蛋白的表达,以34例正常宫颈组织及慢性宫颈炎组织作为对照。比较各组间Notch1及HPV16E6/E7表达的差异,并分析Notch1与HPV16E6/E7表达的关系。结果 Notch1蛋白在细胞胞浆、胞核及胞膜中表达,HPV16E6/E7在细胞核中表达。从对照组到CIN组到宫颈癌组,Notch1及HPV16E6/E7的表达均逐渐增强（P〈0.01）。Notch1的阳性表达与宫颈癌不同分期、分化程度、淋巴结是否转移无关（P〉0.05）。在宫颈癌组中Notch1与HPV16E6/E7的表达均呈正相关性（P〈0.01）。结论 Notch1表达与HPV16E6/E7感染可能与CIN及宫颈癌的发生密切相关,两者在宫颈癌的发病机制中可能协同发挥作用。     

宫颈癌中抑癌基因FHIT表达与HPV16基因状态

目的:探讨抑癌基因FHIT表达及人乳头瘤病毒16(HPV16)的基因型整合状态在宫颈癌发生发展中的作用及相关性。方法:选取柳州市人民医院2013年6月至2014年12月收治的42例宫颈癌、55例宫颈上皮内瘤样病变(CIN)和20例宫颈正常组织的患者,免疫组化法检测宫颈组织中FHIT蛋白表达;多重PCR法检测HPV16 E2/E7表达。结果:FHIT蛋白的总阳性表达率为57.26%(67/117),正常宫颈组织、CINⅠ、CINⅡ、CINⅢ和宫颈癌中FHIT蛋白阳性率分别为85.00%、80.00%、75.00%、60.00%和26.19%。随着宫颈病变加重,FHIT蛋白阳性表达率下降,组间差异有统计学意义(χ~2=7.335;P=0.003)。117例单纯HPV16阳性标本HPV16总整合率为81.20%,正常宫颈组织、CINⅠ、CINⅡ、CINⅢ和宫颈癌中整合率分别为60.00%、66.67%、75.00%、95.00%和92.86%;随病变加重,整合率增强,组间差异有统计学意义(χ~2=5.713,P=0.003);FHIT蛋白阳性表达在不同HPV16整合时不同,差异有统计学意义(χ~2=11.989,P=0.000)。结论:HPV16基因整合可能通过诱导FHIT基因低表达从而促使宫颈癌发生发展。     

HPV16基因整合与FHIT蛋白缺失在宫颈癌变中的作用

目的 探讨HPV16的基因型整合状态及FHIT基因缺失在宫颈癌发生发展中的作用及相关性。方法选取44例宫颈癌、58例宫颈上皮内瘤样病变(CIN)和20例宫颈正常组织的患者,免疫组化检测宫颈FHIT蛋白的表达;并用多重PCR法检测HPV16 E2/E7表达。结果 122例单一HPV16阳性标本HPV16总整合率为78.69%,在正常宫颈组织、CIN1、CIN2、CIN3和宫颈癌中整合率分别为60.00%、66.67%、75.00%、86.96%和88.63%,随病变加重,整合率增强,组间差异有统计学意义(P0.05);FHIT蛋白的总缺失率为43.44%,FHIT蛋白在正常宫颈组织、CIN1、CIN2、CIN3和宫颈癌中的缺失率分别为15.00%、20.00%、25.00%、43.48%和72.73%。FHIT蛋白的缺失率,随病变加重,缺失率增高,组间差异有统计学意义(P0.05)。FHIT蛋白缺失在不同HPV16整合时不同,差异有统计学意义(P0.05)。结论 HPV16基因整合和FHIT蛋白缺失在宫颈癌的发生、发展中起重要作用,HPV16基因整合可能通过诱导FHIT基因低表达从而促使宫颈癌发生发展。     

p16~(INK4A)蛋白及HPV感染与宫颈上皮内瘤变和宫颈鳞癌的关系研究

目的:通过检测慢性宫颈炎(chronic cervicitis)、宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)及宫颈鳞癌(squamous carcinoma of the cervix,SCC)组织中p16INK4A蛋白表达与人乳头瘤病毒(human papillomavirus,HPV)感染的状况,探讨p16INK4A蛋白和HPV感染与宫颈鳞癌发生发展的关系。方法:采用免疫组化PV-9000方法检测114例宫颈组织标本的p16INK4A表达,用核酸分子快速导流杂交基因芯片技术(HybriMax)检测21种HPV DNA。结果:(1)慢性宫颈炎、CINⅠ、CINⅡ、CINⅢ、SCC组织中p16INK4A蛋白表达阳性率分别为0%、45.83%、72.73%、84.00%、91.30%,随着宫颈病变程度加重,p16INK4A蛋白表达阳性率逐渐增高,表达强度增加,差异有统计学意义(P<0.05)。(2)慢性宫颈炎、CINI、CINⅡ、CINⅢ、SCC中HPV感染阳性率分别为20.00%、37.50%、54.55%、56.00%、73.91%,HPV感染在不同宫颈病变组织中差异有统计学意义(P<0.05)。随宫颈病变程度加重,HPV阳性率呈递增趋势,不同病变组织中位于前3位的HPV感染型别分别为:CINⅠ组,HPV16、18、58;CINⅡ～Ⅲ组,HPV16、33、52;宫颈鳞癌组,HPV16、18,52。(3)相关分析结果显示,病变组织中p16INK4A蛋白表达与HPV感染呈正相关(r=0.268,P<0.05)。结论:p16INK4A蛋白表达与宫颈病变程度有关,p16INK4A可能参与了HPV相关的宫颈癌发生。二者联合检测对宫颈癌筛查和预防具有重要意义。     

Distribution of HPV16 and 18 intratypic variants in normal cytology, intraepithelial lesions, and cervical cancer in a Mexican population

OBJECTIVE: Several intratype variants of HPV16 and 18 have been identified. These variants are associated with populations from different geographic regions, and show a differential distribution among the severity of the cervical lesion, most likely due to different pathogenic potential. The objective of this study was to investigate the variant distribution of HPV16 and 18 in a Mexican population and its association with the severity of the cervical lesion and the histological lineage of cervical cancer. METHODS: HPV types 16 and 18 detection was performed in 412 samples of preinvasive and invasive specimens from patients attending a Primary Health-Care Center, an Early Cervical Lesion Clinic, or a Cancer Center. Distribution of HPV variants was correlated with the cytological findings and tumor cell types using contingency tables. Statistical difference was tested with the Fisher's Exact Test or its Fisher-Freeman-Halton extension for RXC tables. Alpha value was set at the P < 0.05. RESULTS: Among the 277 women included in this study without cancer, 63.5% (176 cases) had a normal cytology; from the remaining 101 women, 53.5% were LSIL (54 cases), and 46.5% HSIL (47 cases). From a total of 135 invasive carcinomas, 78.5% were squamous (106 cases); 6.6% adenocarcinoma (9 cases); 9.6% adenosquamous (ADSC) (13 cases); and 5.1% were undifferentiated carcinoma (7 cases). HPV16 E and AA-a were evenly distributed among preinvasive and invasive lesions. However, the isolate AA-c was exclusively found in cervical cancer. HPV18 Var-1(E) was almost exclusively found in invasive lesions, while the HPV18 Var-2(Af) predominated in normal or preinvasive lesions. In invasive cancer, this variant was found only in squamous tumors. CONCLUSIONS: The differential distribution of HPV16 and 18 variants in cervical lesions we found further supports experimental data on the different pathogenic potential of HPV16 and 18 variants for cervical cancer development.     

HPV16/18 prevalence in cervical lesions/cancers and p53 genotypes in cervical cancer patients from India

OBJECTIVES: The HPV16/18 code for an oncoprotein-E6, which binds to p53 tumor suppressor protein and degrades the protein via ubiquitination. A common polymorphism of p53 in exon 4 codon 72, resulting in either proline (Pro) or arginine (Arg), affects HPV16/18 E6-mediated degradation of p53 protein in vivo. Hence, in the current study we investigated the prevalence of HPV16/18 in cervical lesions and the distribution of p53 genotypes in cervical cancers and normal healthy women. METHODS: DNA from 337 Indian women with invasive cervical cancers, 164 women with clinically normal cervix, 64 women with low-grade squamous intraepithelial lesions (LSIL), and 5 women with high-grade squamous intraepithelial lesions (HSIL) was examined for the presence of HPV16/18 using consensus primers in a polymerase chain reaction (PCR), and the specific HPV type was identified by Southern hybridization of the PCR product using HPV16/18 type-specific nucleotide sequences as probes. Further, 134 women with cervical cancers and 131 healthy women were used to determine the frequency of p53 genotypes, Pro/Pro, Arg/Arg, and Pro/Arg, using peripheral blood cell DNA to indicate the constitutional genotypes and allele-specific primers, in a PCR-based assay. RESULTS: We observed a prevalence of HPV16/18 in 77% (258/337) of cervical cancer patients, 38% (24/64) of LSILs, 4 of 5 HSILs, and 15.2% (25/164) of normal healthy women. The frequency of distribution of the three genotypes of p53 codon 72 in a subgroup of the HPV16/18-positive cervical cancer patients was Pro/Pro 0.18 and Arg/Arg 0.26, with the heterozygous Pro/Arg 0.56, differing significantly from the genotype frequency in the normal healthy women (chi(2) = 6.928, df = 2, P < 0.05). CONCLUSIONS: A high prevalence of HPV16/18 was observed in the cervical cancers. The prevalence in LSILs confirms HPV16/18 infection as an early event and further indicates a role in progression of lesions. The p53 genotype distribution indicated that women homozygous for Arg genotype were at a 2.4-fold higher risk for developing HPV16/18-associated cervical carcinomas, compared to those showing heterozygous Pro/Arg genotype (odds ratio 2.4, 95% confidence interval 1.89 to 3.04).     

Aptima法人乳头瘤病毒E6/E7 mRNA检测在子宫颈癌筛查中的流行病学分析及其诊断价值

目的 通过Aptima法检测人乳头瘤病毒(HPV)E6/E7 mRNA,研究其分型和定量检测结果在不同年龄和子宫颈活检中的分布情况,探讨其在子宫颈病变中的诊断价值.方法 选取2019年1月至2020年1月于郑州大学第三附属医院行HPV E6/E7 mRNA检测的患者为研究对象,收集HPVE6/E7 mRNA阳性患者的年...     

HPV L1壳蛋白联合HR-HPV分型、TCT检测对宫颈癌前病变及宫颈癌的诊断价值分析#br#

Objective? To explore the diagnostic value of human papilloma virus L1 (HPV L1) capsid protein combined with high-risk HPV (HR-HPV) typing and thinprep cytology test (TCT) for cervical precancerous lesions and cervical cancer. Methods?1 094 women who received cervical cancer screening were selected as the research subjects, and were grouped based on histopathological results. The expression level of HPV L1 capsid protein, and positive rates of HR-HPV and TCT were compared between the groups. Results?There was a significant difference between TCT positive rate and HR-HPV positive rate (P<0.05). Histopathological examination found 179 normal cases or cases only with inflammation (control group), 26 cases of CIN gradeⅠ (CIN1 group), 44 cases of CIN gradeⅡ (CIN2 group), 40 cases of CIN gradeⅢ (CIN3 group) and 21 cases of cervical cancer (cervical cancer group). The positive rates of TCT and HR-HPV in cervical cancer group were the highest, and the expression level of HPV L1 capsid protein was the lowest (P<0.05). The AUC values of HPV L1 capsid protein combined with HPV typing and TCT to diagnose cervical precancerous lesions and cervical cancer were 0.897 and 0.804, respectively (P<0.05). Conclusion?HPV L1 capsid protein, HPV typing and TCT are of clinical value in the diagnosis of cervical precancerous lesions and cervical cancer.     

青岛地区宫颈病变组织中HPV16 URR和E6基因突变分析

目的:分析青岛地区妇女不同宫颈病变组织中人乳头瘤病毒16型(HPV16)上游调控区(URR)和E6基因序列多态性,以及序列变异与宫颈病变的相关性。方法:采用聚合酶链式反应(PCR)技术检测120例宫颈癌及254例宫颈上皮内瘤变(CIN)标本,筛选出HPV16阳性标本,进而扩增出HPV16URR和E6基因,PCR产物纯化后测序,与德国HPV标准株进行对比分析。结果:URR测序结果发现了19个突变位点,所有标本在7521位点均发生了G→A突变;63.30%(69/109)标本中检测到24,7730和7842的联合突变,为另一突变热点。E6测序结果共发现17个突变位点,突变热点为178位点(D25E),其在宫颈癌组、CINⅢ组和CINⅡ组的比例分别为61.11%(44/72)、62.00%(31/50)和50.00%(16/32),3组的差异无统计学意义(P=0.499)。仅观察到6例350位点突变(L83V)。亚洲型(AS)是最多见的突变类型,在154例HPV16阳性标本中占59.09%,其次为E-P原型(33.12%)。结论:青岛地区妇女宫颈病变组织中HPV16URR突变热点为nt7521以及nt24、nt7730和nt7842的联合突变。HPV16E6突变热点为nt178,这些突变热点可能与癌前病变的进展有密切关系。AS型和原型是青岛地区宫颈病变组织中两种主要的HPV16分支。