HPV、突变型p53和COX-2在外阴HPV感染相关疾病的表达及意义

目的:探讨人乳头瘤病毒(HPV)、突变型p53及COX-2表达与外阴HPV感染疾病的关系及临床意义.方法:应用原位杂交及免疫组化技术,检测57例正常外阴(NCE)、124例外阴尖锐湿疣(VA)、79例外阴上皮内瘤变(VIN)、91例外阴鳞状细胞癌(VSCC)组织中HPV6/ll、HPV16/18、突变型053及COX-2的表达.结果:VA组织中HPV 6/11阳性表达率(60.5％)显著高于VIN、VSCC(30.4％和34.1％)(P＜0.05);VSCC组织中 HPV 16/18阳性表达率(79.1％)显著高于VA、VIN(28.2％和36.7％)(P＜0.05).VA、VIN、VSCC组织中COX-2阳性表达率(62.1％、58.2％和64.8％)均显著高于NCE(10.5％)(P＜0.05);但前3组比较,则无显著差异(P＞0.C5).VA、VIN、VSCC组织中p53表达呈逐渐增高趋势,两两比较均有显著差异(P均＜0.05).NCE组织中HPV 6/11、HPV16/18及突变型p53均呈阴性表达.结论:外阴活检时行HPV 6/11、HPV16/18、突变型p53及COX-2检测,有助于外阴恶性病变的早期诊断.     

人乳头瘤病毒及bcl-2的表达与维吾尔族外阴癌发病关系的研究

目的:探讨人乳头瘤病毒(human papilloma virus,HPV)感染及bcl-2蛋白的表达在维吾尔族妇女外阴癌发生发展中的作用及其相互关系。方法:应用原位杂交及免疫组化SP法对67份维吾尔族妇女外阴手术切除存档蜡块(其中外阴癌57份,癌旁正常外阴组织10份)进行HPV16/18型及bcl-2蛋白的检测。结果:(1)57份外阴癌及10份癌旁正常外阴组织中,无一例HPV16/18型阳性表达;bcl-2蛋白在外阴癌组织中阳性表达率为70.18%(40/57),在10份癌旁组织中表达均阴性。(2)bcl-2在Ⅰ期、Ⅱ期及Ⅲ~IV期外阴癌中分别为70.18%,75.0%及50.0%,其在各期别之间无显著性差异(P=0.302),bcl-2蛋白表达在外阴癌淋巴结阳性组(88.88%)明显高于淋巴结阴性组(51.62%),两者之间有显著性差异(P=0.044)。结论:维吾尔族外阴角化型鳞状上皮癌的发生可能与HPV感染无关。bcl-2的表达与维吾尔族外阴角化型鳞状上皮癌密切相关。bcl-2的表达与外阴癌临床分期及组织学分级无明显相关,但与淋巴结转移有关。     

基质金属蛋白酶MMP-9及其抑制剂TIMP-1在外阴癌中的表达

目的　观察基质金属蛋白酶MMP 9和其抑制剂TIMP 1在外阴癌组织中的表达情况 ,探讨其与外阴癌的浸润及转移的关系。方法　采用免疫组织化学SP方法对 33例外阴癌、2 3例VIN、15例外阴色素减退疾病、14例外阴尖锐湿疣和 12例正常皮肤组织进行标记及分析。结果　MMP 9、TIMP 1蛋白在以上 5组细胞浆中的阳性表达分别为 31/33、2 4 /33,2 1/2 3、19/2 3,15 /15、13/15 ,12 /14、12 /14 ,10 /12、7/12。MMP 9、TIMP 1蛋白在外阴癌、VIN、色素减退疾病、尖锐湿疣中的阳性表达率与正常对照组相比差异均无显著性意义 (P >0 0 5 ) ,但在(-～ +)组与 (++～ +++)组表达率之间MMP 9在外阴癌组与其他各组间相比差异均有显著性意义 (P<0 0 5 ) ;TIMP 1在外阴癌组与色素减退疾病组和正常组相比差异有显著性意义 (P <0 0 5 )。这 2个指标在外阴癌中的表达与临床病理特征无明显相关性。在外阴癌中MMP 9与TIMP 1在阳性表达之间差异有显著性意义 (P <0 0 5 ) ,而强阳性表达间差异有极显著性意义 (P <0 0 1)。结论　在外阴癌中MMP 9的表达强度超过TIMP 1时可能是外阴癌侵袭的一个早期指标 ,但不能单独作为预测外阴癌结局的一个标志。     

HPV16 18 E6蛋白在宫颈脱落细胞中的表达研究

目的 探讨人乳头状瘤病毒(human papillomavirus,HPV)16和(或)18 E6蛋白在宫颈脱落细胞中的表达与宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)及宫颈鳞状细胞癌(cervical squamous cell carcinoma,CSCC)的关系.方法 采用免疫印迹(Western blot)方法检测2003年6月至2005年6月中国医科大学第一附属医院CSCC 30例、CIN 52例和16例正常宫颈脱落细胞中HPV16和(或)18 E6蛋白的表达.结果 在上述各组宫颈脱落细胞中HPV16和(或)18 E6表达阳性率分别76.7%(23/30)、51.9%(27/52)和12.5%(2/16);其中在CSCC宫颈脱落细胞中HPV16和(或)18 E6明显高于正常对照组的表达(P＜0.01);CINⅡ、Ⅲ组的表达也显著高于对照组(P＜0.05),而CINⅠ组与正常对照组比较差异无显著性意义(P＞0.05).结论 宫颈脱落细胞标本存在HPV16和(或)18 E6癌基因的表达.宫颈脱落细胞中HPV16和(或)18 E6癌基因的过度表达与CSCC及CINⅡ、Ⅲ密切相关.     

EGF与外阴癌发生发展的关系

目的:通过检测表皮生长因子(EGF)在正常外阴皮肤、外阴上皮内瘤变(VIN)和外阴鳞状细胞癌(VSCC)中的表达,探讨其在外阴癌变中的作用。方法:应用免疫组化SP法进行检测。EGF的表达与外阴肿瘤的临床病理特征相比较。结果:正常外阴组织、低级别VINs、高级别VINs、外阴癌中EGF蛋白阳性表达率分别为0(0/20)、18.2%(2/11)、82.8%(9/11)、100%(60/60),P<0.05。在高、中、低分化的浸润性外阴癌中,EGF的细胞阳性表达率分别为36.2%、42.7%、47.1%,P>0.05。有淋巴结转移者EGF蛋白阳性表达率为30.2%明显低于无淋巴结转移者的49.8%。结论:EGF蛋白的异常表达在外阴癌的发生发展中起着重要的作用,而且可为外阴癌的预后提供重要的信息。     

外阴病变组织中精子相关抗原-9的表达及意义

目的:研究外阴鳞状上皮内瘤变(VIN)和外阴鳞状细胞癌组织中精子相关抗原-9(SPAG9)的表达及其潜在的临床意义。方法:应用免疫组织化学技术检测16例VIN、41例侵袭性外阴鳞状细胞癌和10例正常外阴皮肤组织中SPAG9蛋白的表达。结果:SPAG9蛋白在VIN及侵袭性外阴鳞状细胞癌中的表达率均为100%,在正常外阴皮肤中表达率为0;根据H-score分值比较:SPAG9蛋白在外阴鳞状细胞癌组织中的表达高于VIN组织,G2～G3肿瘤组织中SPAG9蛋白表达高于G1者,有淋巴结转移者肿瘤组织内SPAG9蛋白的表达高于无淋巴结转移者,差异均有统计学意义(P<0.05);外阴鳞状细胞癌Ⅱ～Ⅲ期患者肿瘤组织中SPAG9蛋白的表达高于Ⅰ期,差异无统计学意义(P>0.05)。结论:SPAG9蛋白可能在外阴癌的发生和发展发挥重要作用,有望成为外阴癌早期诊断和预后评估的生物学标记物。     

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.     

p27蛋白在子宫内膜癌组织中的表达及其临床意义

目的 :探讨p2 7在子宫内膜癌组织中的表达及其临床意义。方法 :采用免疫组化S -P法测定 16份正常子宫内膜、18份子宫内膜不典型增生及 50份子宫内膜癌组织中的p2 7蛋白表达。结果 :p2 7蛋白在子宫内膜癌、子宫内膜不典型增生及正常子宫内膜中的表达率分别为 34%、6 6 .6 7%和 93.75% ,正常子宫内膜及不典型增生组均显著高于子宫内膜癌组 ,差异有显著性 (P <0 .0 5)。p2 7蛋白表达与子宫内膜癌组织学分级、肌层浸润程度及患者预后显著相关 (P <0 .0 5) ,但与临床分期无关。结论 :p2 7蛋白表达下降或缺失可能在子宫内膜癌的发生、发展中起重要作用 ,并可能提示患者的预后     

cyclinD1、p16蛋白在子宫内膜癌中的表达及其临床意义

目的 :探讨cyclinD1、p16与子宫内膜癌发生与发展的关系。方法 :用免疫组化SP法检测了 50例子宫内膜癌、2 2例子宫内膜不典型增生、10例正常子宫内膜组织中cyclinD1、p16蛋白的表达情况。结果 :cyclinD1蛋白在正常子宫内膜、内膜不典型增生、子宫内膜癌中的表达率分别为 0 ( 0 /10 )、2 2 .73% ( 5/2 2 )、4 0 .0 0 % ( 2 0 /50 ) ,其中子宫内膜癌与正常内膜差异有显著性 (P <0 .0 5)。p16蛋白在内膜癌的表达率为 4 6.0 0 % ,明显低于正常内膜组织的 90 .0 0 % (P <0 .0 5)。 50例内膜癌中 ,cyclinD1蛋白在G2 、G3级及临床Ⅱ～Ⅲ期中的表达率分别高于G1级及临床Ⅰ期 (P <0 .0 5) ,复发组表达率高于未复发组 (P <0 .0 5)。p16蛋白表达与子宫内膜癌的细胞分级、临床分期及预后有关 (P <0 .0 5) ,p16蛋白表达阴性者分化差 ,分期晚 ,复发率高。相关性分析显示 ,cyclinD1与p16表达呈负相关 ,二者协同异常表达者生物学行为较差。结论 :cyclinD1、p16与子宫内膜癌的发生、发展密切相关 ,二者异常表达提示子宫内膜癌复发率高、预后差     

宫颈癌组织中HPV、EBV、p53、bcl-2基因产物表达及临床病理意义

目的 :探讨人乳头瘤病毒 (HPV)、EB病毒 (EBV)和p5 3、抗细胞凋亡因子 (bcl- 2 )基因产物在宫颈癌中的表达及临床意义。方法 :对 2 0例正常宫颈组织标本及 5 0例宫颈癌标本石蜡包埋制成切片 ,采用免疫组织化学染色方法(LSAB)检测HPV、EBV产物和p5 3、bcl 2、基因产物表达。结果 :在宫颈癌组织中HPV、EBV的阳性表达率为 4 4 %、5 2 % ,明显高于正常宫颈组织的 5 %、10 % (P <0 .0 5 ) ,HPV和EBV的混合感染率为 18% (9/ 5 0 )。p5 3基因产物阳性表达率宫颈癌组 (2 4 % )高于正常宫颈组织 (0 % ) (P <0 .0 5 )。在宫颈癌组织中bcl 2基因产物的阳性表达率 (72 % )明显高于正常宫颈组织的 (2 0 % ) (P <0 .0 5 )。HPV与p5 3有相关性 ,EBV与bcl 2相关。结论 :①HPV、EBV在宫颈癌组织中的感染率明显高于正常人。②HPV可能通过造成p5 3基因产生突变而失活 ,导致宫颈癌的发生 ;EBV可能通过使bcl 2基因蛋白产生上调性表达 ,抑制细胞凋亡 ,导致宫颈细胞癌变     

Immunohistochemical staining for p16 and p53 in premalignant and malignant epithelial lesions of the vulva.

Two distinct types of vulvar squamous cell carcinomas and their precursors, vulvar intraepithelial neoplasias (VIN), which differ in terms of clinical presentation and behavior, have been delineated. Human papillomavirus (HPV)-associated carcinomas are of basaloid or warty type, whereas tumors unrelated to HPV are usually keratinizing and differentiated. Thus, the major stratifying factor for vulvar carcinomas and VIN is their etiopathogenetic relationship with HPV. However, because of technical difficulties in confidently detecting HPV in tissues, this diagnosis is usually based on purely morphologic criteria, even though some overlap exists between these histologic types. Recently, the tumor suppressor protein p16 has been shown to be specifically overexpressed in HPV-related carcinomas and premalignant lesions of the uterine cervix, oral cavity, and anus, but the presence of p16 vulvar squamous lesions has not been examined. We have evaluated the immunohistochemical expression of p16 in a series of formalin-fixed, paraffin-embedded vulvar carcinomas and their putative precursors. p16 was strongly positive in all cases of basaloid/condylomatous VIN3 (30/30) and basaloid (7/7) and warty (3/3) carcinomas. In contrast, p16 was almost consistently negative in normal skin, squamous cell hyperplasia (0/20), lichen sclerosus (0/19), differentiated (simplex) VIN3 (0/11), verrucous carcinoma (0/2), and keratinizing squamous cell carcinoma (3/33, 9%). One of the keratinizing squamous cell carcinomas positive for p16 occurred in a 25-year-old woman and the other two were associated with small foci of basaloid VIN3 adjacent to the tumor, suggesting a probable relationship with HPV. p16 was positive in 6 of 10 of basal cell carcinomas. In conclusion, p16 immunostaining is a good discriminator between HPV-associated and HPV-unrelated vulvar carcinomas and VIN, although it cannot differentiate basaloid squamous and basal cell carcinoma.     

Human papillomavirus, Epstein-Barr virus and p53 mutation in vulvar intraepithelial neoplasia

OBJECTIVE: To clarify the role of human papillomavirus (HPV) and Epstein-Barr virus (EBV) infection in vulvar carcinogenesis in relation to the mutated p53 gene. STUDY DESIGN: Polymerase chain reaction (PCR) was used to amplify DNA sequences of the viruses and PCR-single-strand conformation polymorphism analysis to screen for p53 gene mutations in exons 5-8 from formalin-fixed, paraffin-embedded blocks including 10 undifferentiated vulvar intraepithelial neoplasia (VIN) specimens. RESULTS: HPV and EBV DNA was found in 75% (6/8) and 0% (0/10) of VIN tissues, respectively. Oncogenic HPV 16 was the predominant type. HPV DNA extraction was not possible in 2 VIN specimens. p53 Gene mutation was shown in 20% (2/10) of VIN lesions. No correlation was found between p53 gene mutation the presence of viral HPV or EBV DNA. Mutated p53 was equally distributed between HPV-positive and -negative VIN cases. CONCLUSION: Our results suggest that although most undifferentiated VIN lesions are associated with HPV infection, p53 mutations may occur independent of viral infection even in the presence of oncogenic HPV. HPV, but not EBV or p53 gene mutation, can play a role in the pathogenesis of undifferentiated VIN.     

Probable nonpapillomavirus etiology of squamous cell carcinoma of the vulva in older women: a clinicopathologic study using in situ hybridization and polymerase chain reaction.

A clinical, pathologic, and molecular virologic analysis of 30 cases of invasive squamous cell carcinoma of the vulva was undertaken to investigate the relationship of human papillomavirus (HPV) to this neoplasm. The presence of the virus was detected by the polymerase chain reaction and localized in the tumor and in the adjacent epithelium by in situ hybridization of paraffin sections of vulvectomy specimens. Specimens were examined for nucleic acid sequences of HPVs 6, 11, 16, and 18 were detected by in situ hybridization utilizing 35S-labeled antisense RNA probes and by polymerase chain reaction using HPV type-specific primers for a segment of the E6 gene followed by Southern hybridization of the amplified products. The cases were classified as typical squamous cell carcinoma, basaloid carcinoma, and warty carcinoma. Typical squamous cell carcinoma shows varying degrees of squamous maturation, whereas basaloid carcinoma is characterized by immature basal-type cells showing minimal or no squamous maturation. Warty carcinoma displays an exophytic condylomatous appearance. The squamous cells of this tumor are mature, and many show koilocytotic atypia characterized by a variable degree of nuclear atypia and cytoplasmic vacuolization. The adjacent epithelium was classified as squamous hyperplasia, lichen sclerosus, or vulvar intraepithelial neoplasia (VIN). VIN was subdivided into basaloid or warty VIN using similar criteria as for the invasive carcinomas. Overall, HPV 16 was detected in 11 cases and HPV 18 in two; none of the cases were positive for HPVs 6/11. HPV was detected in four (21%) of 19 squamous cell carcinomas, six (75%) of eight basaloid carcinomas, and three (100%) of three warty carcinomas. The adjacent epithelial lesions also showed a close correlation with the tumor type and presence of HPV. Fourteen (74%) squamous cell carcinomas had adjacent squamous hyperplasia; all of these squamous hyperplasias were negative for HPV. In contrast, seven (87%) of the basaloid carcinomas had adjacent basaloid-VIN and HPV 16 was detected within the VIN in three. Three warty carcinomas (100%) had adjacent warty VIN or basaloid VIN, and HPV was detected within VIN in two. The mean age of women with squamous cell carcinoma was 77 years, for women with basaloid carcinoma 54 years, and for those with warty carcinoma 47 years. The mean age of women with HPV-negative tumors was 77 years compared with 55 years for women with HPV-positive tumors (p less than 0.01). Thus, there appears to be a close correlation between the presence of HPV, specific subsets of invasive carcinoma and VIN, and age.(ABSTRACT TRUNCATED AT 400 WORDS)     

Search for herpes simplex virus 2 and human papillomavirus genetic expression in vulvar neoplasia

Specimens from vulvar carcinomas and vulvar intraepithelial neoplasia (VIN) of various degrees were analyzed for the presence of herpes simplex virus 2 (HSV 2) and human papillomavirus (HPV) genetic information. A search for the HPV 16 E6 protein as well as the HSV 2 antigenic determinant LA1 and ICSP 11/12 protein was carried out with an immunoperoxidase assay on 12 vulvar carcinomas and 6 VINs. Seven invasive cancers and four VINs were screened for the presence of homology to HPV 16 DNA and HSV 2 DNA transforming sequences with Southern blot hybridization. We used specimens from labial tumors and from normal vulvas and cervixes as controls. The preliminary results showed that one vulvar carcinoma and two VINs contained HPV 16 DNA. Four vulvar carcinomas expressed the E6 protein, while all the VINs were negative. Homology to HSV 2 DNA transforming sequences was detected in one vulvar cancer but not in any VIN cases. Positivity to HSV 2 ICSP 11/12 was observed in 33.3% of VIN cases and 75% of invasive cancers. HSV 2 LA1 antigenic determinant was expressed in 33.3% of VIN and 66.6% of cancer cases.     

子宫颈癌p53抑癌基因突变与病毒感染的研究

采用聚合酶链反应－单链构象多态性分析（ＰＣＲ－ＳＳＣＰ）、多重引物ＰＣＲ和巢式引物ＰＣＲ，对同一宫颈癌组织中ｐ５３基因（第６～８外显子）突变以及人乳头状瘤病毒（ＨＰＶ）和人巨细胞病毒（ＨＣＭＶ）感染进行相关性研究。并与正常宫颈组织进行对照。结果：３８例宫颈癌组中，２例有ｐ５３第７外显子突变。其中１例伴有ＨＰＶ１６感染，２例伴有ＨＣＭＶ感染。宫颈癌组ＨＰＶＩ６、１８ＤＮＡ的阳性率为６３．２％（２４／３８），ＨＣＭＶＤＮＡ为８４．２％（３２／３８）。２１例对照组中，ＨＰＶ１６、１８ＤＮＡ和ＨＣＭＶＤＮＡ的阳性率分别为４．８％和３８．１％（Ｐ＜０．００５）。ＨＰＶ１６、１８阳性的子宫颈癌中，８７．５％伴有ＨＣＭＶ感染。对照组中，无一例同时检测出ＨＰＶ１６、１８和ＨＣＭＶ者。提示：宫颈癌组织中，ｐ５３基因突变并不常见，其突变与ＨＰＶ１６、１８感染无显著关系。宫颈癌与ＨＰＶ１６、１８关系密切，ＨＣＭＶ可能与ＨＰＶ协同作用，导致宫颈癌的发生。     

In situ hybridization analysis of human papillomavirus DNA segregation patterns in lesions of the female genital tract

Various histologic features may be used to divide human papillomavirus (HPV)-related lesions of the genital tract into two groups: condylomata and "low-grade" or grade 1 cervical intraepithelial neoplasias (CIN 1) versus "high-grade" or grade 2 and 3 intraepithelial neoplasias. Using in situ hybridization analysis we correlated HPV DNA type with histologic features in 350 biopsies of lesions from the cervix, vulva, and perianal region. HPV DNA was most commonly found in vulvar and perianal condylomata (39/46, 85%), whereas the rate in CIN 1 lesions was 72% (86/120). The rates were 53% (40/76) and 57% (12/21) in CIN 2/3 and vulvar intraepithelial neoplasm (VIN) grades 2 and 3, respectively. The HPV type in all but 2 of the 39 perianal and vulvar condylomata which contained HPV was 6/11. Despite their similar histologic features, the HPV type in only 23 of 86 (27%) CIN 1 cases with detectable HPV was 6/11 compared to 31 of 86 (36%) which contained HPV 16-related DNA and 32 of 86 (37%) which contained HPV 31,-33, or -35-related DNA. The viral DNA in the majority of CIN 2/3 lesions and all of the VIN 2/3 lesions was HPV-16 related; no CIN 2/3 or VIN 2/3 lesion had HPV 6/11-related DNA. It is concluded that although cutaneous genital tract condylomata are highly associated with HPVs of low oncogenic potential (types 6 and 11), these HPV types are not as frequent as the oncogenic HPVs (16, 31, 33, and 35) in CIN 1 lesions. Further, HPV 6/11 appears to be very rarely associated with CIN 2/3 or VIN 2/3 lesions.     

Genetic changes during the multistage pathogenesis of human papillomavirus positive and negative vulvar carcinomas.

OBJECTIVE: To identify the molecular alterations found in 30 human papillomavirus (HPV) positive (n = 15) and negative (n = 15) vulvar carcinomas (VC) and their associated preinvasive lesions (VIN [vulvar intraepithelial neoplasia]) and normal epithelium to determine a common molecular pathogenesis of HPV positive and negative VC. METHODS: Loss of heterozygosity (LOH) at seven 3p chromosomal regions (3p12, 3p14.2, 3p14.3-21.1, 3p21.3, 3p22-24, 3p24.3, 3p25), 13q14 (RB) and 17p13.1 (p53) loci, and TP53 gene mutations in microdissected archival tissues were investigated. RESULTS: Fourteen of fifteen HPV positive VC had HPV 16 DNA sequences. The fractional regional loss index (FRL), an index of total allelic loss at all chromosomal regions analyzed, was greater in the HPV negative VCs than in the HPV positive tumors (FRL = 0.55 versus 0.32; P = .048) and was also greater in the HPV negative high-grade VINs as compared with the HPV positive lesions (0.29 versus 0.02; P = .002). Overall, LOH at any 3p region was frequent (80%) in both groups of cancers and in their associated VIN lesions. Although TP53 gene mutations were present in a minority of VCs (20%), allelic losses at the TP53 locus were frequently present, especially in HPV negative VCs, as compared with the HPV positive tumors (62% versus 15%; P = .02). CONCLUSION: A greater number of molecular alterations are found in HPV negative VCs compared with HPV positive tumors. Allelic losses at 3p are common early events in vulvar carcinogenesis in HPV negative cancers detected at a high rate in the corresponding high-grade precursor lesions (VIN II/III). TP53 gene mutations with associated 17p13.1 LOH are more common in HPV negative cancers.     

Vulvar squamous cell carcinoma is a multifactorial disease following two separate and independent pathways.

Two separate pathways leading to vulvar carcinoma have been suggested. First, a human papillomavirus (HPV)-dependent pathway, in which premalignant stages of vulvar cancer are the classic vulvar intraepithelial neoplasia (VIN) lesions. Second, an HPV-independent pathway, associated with differentiated VIN III lesions and/or lichen sclerosus. To obtain insight into the mechanisms underlying these pathways, we determined the relationship between HPV DNA and the expression of p14(ARF) and p16(INK4A) in non- and (pre)malignant vulvar lesions. Seventy-three archival samples of non- and (pre)neoplastic vulvar lesions were selected and tested for hr-HPV DNA using a broad-spectrum HPV detection/genotyping assay (SPF(10)-LiPA) and the expression of p14(ARF) and p16(INK4A). The prevalence of HPV increased with the severity of the classic VIN lesions; in VIN I no hr-HPV was detected, in VIN II 43%, and in VIN III 71% of the samples were hr-HPV-positive. Roughly the same was true for the expression of p14(ARF) and p16(INK4A). The simultaneous expression of p14(ARF) and p16(INK4A) was highly associated with the presence of hr-HPV DNA. Hr-HPV was detected in only a single case of the differentiated VIN III lesions, whereas no expression of p14(ARF) was found and 16(INK4A) was present in only two cases. All 16 samples of vulvar cancer were hr-HPV DNA- negative, although in respectively 63% and 25%, p14(ARF) and p16(INK4A) was expressed. No relation was found between hr-HPV and the expression of p14(ARF) and p16(INK4A) in the 20 nonneoplastic vulvar lesions. Our results provide further evidence that vulvar squamous cell carcinoma is a multifactorial disease that develops from two different pathways. First, an HPV-dependent pathway with a remarkable resemblance to CIN lesions and cervical carcinoma and second, an HPV-independent pathway in which differentiated VIN III lesions that are hr-HPV-negative may be precursors.     

Vulvar intraepithelial neoplasia p53 expression, p53 gene mutation and HPV in recurrent/progressive cases

OBJECTIVE: To evaluate p53 protein overexpression and p53 gene mutation in primary and recurrent undifferentiated vulvar intraepithelial neoplasia (VIN), establishing the recurrence and progression rates, median time interval, and sites of the initial lesion and first recurrence, addressing the relationship with HPV infection. STUDY DESIGN: Twenty women with undifferentiated VIN treated with wide surgical excision were followed every 6 months for 7 years and divided into groups with and without recurrence/progression. p53 Protein was detected in paraffin sections using the monoclonal p53 antibody. DNA was extracted from paraffin sections. Polymerase chain reaction/single strand conformation polymorphism (PCR-SSCP) analysis was utilized to screen for p53 gene mutations in exons 5-8. HPV was determined by digesting PCR products with restriction endonucleases. RESULTS: Recurrences were observed in 8 (40%) patients and progression to cancer in 1 (5%). Two cases recurred twice. The median interval for recurrence/progression was 24.5 months. Recurrent/progressive lesions were located in the same area of the initial lesions in 10 cases (91%). p53 Overexpression was observed in 50% (10/20) of primary lesions, of which 45% corresponded to the 9 recurrent/progressive cases. p53 Overexpression was detected in 81.8% (9/11) of recurrent/progressive cases. In the last 2 cases PCR-SSCP showed p53 gene mutation. The rate of HPV infection was higher in the group without recurrence. CONCLUSION: p53 Gene mutation plays an important role in undifferentiated VIN pathogenesis independent of high-risk HPV infection and may predict recurrence or progression to vulvar cancer. Undifferentiated VIN recurrent/progressive VIN lesions have a tendency to occur in the same area of the initial lesions, suggesting a molecular disturbance.