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.     

Immunohistochemical examination oncogenic c-erb-b2, egf-r proteins and antioncogenic p53 protein in vulvar cancers HPV-16 positive and negative

In vulvar cancers HPV 16 positive, HPV 16 negative, and vulvar and vaginal precancerous status (VIN, VAIN) immunohistochemical technik onko- and antioncogenic proteins were evaluated. The contrary correlation between HPV 16 presence and overexpression p53 were detected. It suggest the heterogenic etiology of these cancers. There was payed attention to high activity of virus replication and intensive virion production in VAIN. Nontreated vaginal lesion may make difficult to obtain a positive cervical HPV infection treatment.     

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.     

Expression of topoisomerase IIalpha,Ki-67, proliferating cell nuclear antigen,p53, and argyrophilic nucleolar organizer regions in vulvar squamous lesions

OBJECTIVE: It was the aim of this study to investigate the expression of topoisomerase IIalpha (topo IIalpha), Ki-67, proliferating cell nuclear antigen (PCNA), p53, and argyrophilic nucleolar organizer region (AgNOR) staining in normal vulvar epithelia (NE, N = 10), vulvar condylomas (VC, N = 24), vulvar intraepithelial neoplasia (VIN, N = 26), as well as squamous cell carcinomas (SCC, N = 22) of the vulva. METHODS: Formalin-fixed, paraffin-embedded archival tissue sections were immunostained with monoclonal antibodies against topo IIalpha, p53, and PCNA, as well as an affinity-isolated prediluted ready-to-use Ki-67 antibody using a standard immunohistochemical method, and stained with a colloid silver solution for AgNORs. Immunostaining was quantitated by determining the percentage of positively staining nuclei in each sample to express the labeling indices (LIs) by counting the immunoreactive nuclei in 1000 epithelial cells per case for each antibody. In each specimen 200 nuclei were examined using a x100 oil emersion lens, and the mean number of AgNORs per nucleus (AC) was calculated. RESULTS: The LIs for topo IIalpha, Ki-67, and PCNA as well as ACs increased stepwise from NE to VCs, VIN lesions, and SCCs. In contrast to PCNA LIs and ACs, a consistent correlation in all four groups was found for Ki-67 and topo IIalpha, suggesting that the latter is a proliferation-associated marker in these tissues. p53 expression was seen 8.3% of VCs, 30.8% of VIN lesions, and 54.45% of SCCs. p53 LIs were not correlated with LIs for topo IIalpha or Ki-67 in SCCs. The LIs for topo IIalpha, Ki-67, PCNA, p53, and ACs were not related to tumor progression, FIGO stage, or tumor grade in SCCs. CONCLUSIONS: This study presents topo IIalpha and Ki-67 as useful proliferation-associated markers of vulvar epithelia.     

外阴癌中HPV感染与p53、MDM2表达的关系

目的 :探讨HPV6/ 11、16/ 18在外阴癌组织中的感染情况及与p5 3、MDM2蛋白表达的关系。方法 :用原位杂交法 (ISH)检测HPV6/ 11、16/ 18在 30例外阴癌、2 1例外阴上皮内瘤变 (VIN)及 10例外阴正常皮肤组织中的表达。同时用免疫组化SP法检测p5 3、MDM2蛋白的表达。结果 :HPV6/ 11、16/ 18在外阴癌、VIN中的阳性表达率分别为 60 % (18/30 )、33.33% (10 / 30 ) ,4 2 .86% (9/ 2 1)、2 8.5 7% (6/ 2 1) ,正常对照组没有表达。p5 3、MDM2蛋白在外阴癌、VIN、正常对照组中的阳性表达率分别为 63.33% (19/ 30 )、4 0 .0 0 % (12 / 30 ) ,4 7.62 %(10 / 2 1)、5 2 .38% (11/ 2 1) ,0 % (0 / 10 )、0 % (0 / 10 )。HPV6/ 11的表达在外阴癌组、VIN组与正常组差异有显著性 (P <0 .0 5 ) ,外阴癌组HPV16/ 18表达与正常组差异有显著性 (P <0 .0 5 )。外阴病变各组p5 3、MDM2与正常组差异有显著性 (P <0 .0 5 )。结论 :HPV6/ 11、HPV16/ 18、p5 3、MDM2蛋白在外阴组织的不同病变中表达均差异有显著性 ,在外阴癌的发生发展中HPV感染、p5 3突变和MDM2表达可能起一定的作用     

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.     

Galectin-3 and CD1a-positive dendritic cells are involved in the development of an invasive phenotype in vulvar squamous lesions.

In this study, the expression patterns of Galectin-3 (Gal-3) and the frequency of infiltrating CD1a positive dendritic cells (DCs) were determined in 82 cases of vulvar tissues, consisting of normal squamous epithelia (NE, N = 10), vulvar condylomas (VC, N = 24), high grade vulvar intraepithelial neoplasias (HG-VIN, N = 26) of common type, and invasive keratinizing squamous cell carcinomas (SCC, N = 22) by a standard immunohistochemical method using monoclonal antibodies to investigate their differential expression in vulvar squamous dysplasia and infiltrating carcinomas with an emphasis on neoplastic transformation and progression. Gal-3 expression was cytoplasmic, nuclear or membranous in NE, VCs, and HG-VINs, with negative or weak and occasionally moderate reactivities. In SCCs, exclusively cytoplasmic staining patterns with moderate or strong reactivity in 59% of cases were observed (p < 0.0001, chi-square test); Gal-3 expression was not related with stage, grade, and recurrence. The frequency of CD1a positive DCs increased from NE and VCs to highest numbers in HG-VINs, was lowest in SCCs (p < 0.0001, ANOVA), and was not related with stage and grade, but with recurrence in SCCs (p = 0,048, t-test). This study indicates that qualitative and quantitative changes of Gal-3 immunoexpression and infiltration by CD1a positive DCs in vulvar NE, VCs, and HG-VIN lesions, respectively, compared with SCCs play a role in the development of an infiltrative phenotype, and may provide adjunctive criteria in the diagnosis of invasion of vulvar squamous epithelia.     

Epidemiology of vulvar intra-epithelial neoplasias

The vulvar intraepithelial neoplasia has been identified as one of the 12 neoplasias whose incidence increases in the developed countries. The vulvar intraepithelial neoplasia (VIN) and invasive vulvar cancer incidence increases by 2.4% per annum; and this principally in young women. The VIN account for 57% of the vulvar neoplasias and are actually more frequent than invasive carcinomas. In the United States, between 1973 and 2000, the incidence of the VIN increased by 411% against 20% for invasive cancers. Similar figures were reported from Norwegian registers. The VIN have a different age distribution than invasive cancers: the incidence of the VIN increases until the age of 40-49 years then decreases while the incidence of invasive cancers increases after 50 years without real peak of incidence. The increase in the incidence of VIN could be followed by an increase in the incidence of invasive cancers but the unknowns on the natural history of the VIN and the impact of the treatments make any extrapolation hazardous. The association between the VIN and the human papillomavirus (HPV) has been well established. It should be noted that, contrary to the cervical neoplasia that are related for nearly 100% to the HPV, only 30-40% of invasive cancers of the vulva are related to HPV, while the other carcinomas are related to the evolution of a vulvar lichen sclerous. The HPV induce various types of anogenital lesion according to their genotype. These lesions can be benign for the HPV6 and 11 and preneoplastic or neoplastic for the HPV16 and 18. The presence of HPV16 and 18 is found in 70 to 80% of the VIN suggesting that HPV vaccines could decrease the incidence VIN and HPV related invasive vulvar cancer.     

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.     

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.     

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.     

PTEN mutation, expression and LOH at its locus in ovarian carcinomas. Relation to TP53, K-RAS and BRCA1 mutations

OBJECTIVE: We aimed to evaluate frequency of PTEN mutation, LOH and expression in ovarian tumors. In search for a molecular pathway, we confronted PTEN gene mutations with TP53, K-RAS and BRCA1 gene status in the same tumors. We also evaluated clinical significance of PTEN expression in a subgroup of patients uniformly treated with platinum-based regimens. METHODS: Molecular analysis was performed on 105 ovarian tumors (100 carcinomas) with the use of the SSCP and sequencing. Seventy-six tumors were analyzed for LOH at 10q23 locus with the use of six polymorphic markers. Immunohistochemical PTEN expression was done on paraffin-embedded material. Multivariate and univariate analysis was performed with the STATA program. RESULTS: PTEN mutations occurred in 5/100 (5%) of all carcinomas and in 3/15 (20%) of endometrioid carcinomas (EC). Low-grade EC that developed in borderline tumors had PTEN and/or K-RAS mutation (4/5, 80%), while high-grade EC had TP53 mutations only. There was a reverse association between PTEN and TP53 mutations (P = 0.005). LOH at PTEN locus was found in 60% of endometrioid and in 28% of serous and clear cell carcinomas. PTEN expression did not associate with PTEN mutations or LOH. Strong PTEN expression diminished risk of death in a TP53 positive group only (HR = 0.35, P = 0.029). CONCLUSION: Our results suggest that PTEN mutations may play a role in a development of low-grade endometrioid tumors. PTEN haploinsufficiency caused by LOH or epigenetic events may possibly contribute to development of other histological types and may be an adverse prognostic factor.     

Vulvar squamous cell carcinoma in young women: a clinicopathologic study of 21 cases.

OBJECTIVES: Invasive squamous cell carcinoma (ISCC) of the vulva occurs most often in older women and the clinical, pathological, and immunohistochemical features of vulvar ISCC in young women are poorly characterized. The aim of this study was to examine clinical and pathological features of ISCC presenting in women younger than 40 years of age. METHODS: Patients younger than 40 years of age who presented with vulvar ISCC were identified in the population-based tumor registry of the British Columbia Cancer Agency (BCCA) for the period 1970-1998. Clinical data and follow-up were obtained. The pathologic material was reviewed and morphologic features assessed. Immunohistochemical staining for MIB-1 and p53 proteins was done and the presence of human papillomavirus (HPV) DNA was assessed by microdissection/PCR. RESULTS: Twenty-one cases, accounting for 5% of all cases of vulvar ISCC encountered at BCCA during this period, were identified, with patient's ages ranging from 17 to 39 years (mean 33). The number of cases of vulvar ISCC in young women, as a percentage of all cases of vulvar ISCC, increased significantly over the study period. Lichen sclerosus was seen in 3 cases. Vulvar intraepithelial neoplasia (VIN) was present in 20 of 21 cases and was multifocal in 4 of them. VIN was subclassified as warty in 7 cases, mixed warty and basaloid in 6, basaloid in 4, and differentiated in 3. There was MIB-1 immunostaining throughout the full thickness of warty and basaloid VIN. Only basal cells stained for MIB-1 in differentiated VIN. Increased p53 expression was present in only 2 cases; both were differentiated-type VIN. HPV DNA was detected in 17 of 20 cases. The tumors were staged as follows: stage IA, 3 cases; stage IB, 13 cases; stage II, 3 cases; stage III, 2 cases. Depth of invasion ranged from <1 to 8.5 mm. The definitive surgical procedure was vulvectomy with lymph node dissection in 14 cases, wide local excision in 6, and excisional biopsy in 1. Clinical follow-up of 1 to 28 years (median, 5 years) showed that 5 patients had local recurrence and 2 died of disease. Of the 21 patients in this study, 1 had concurrent HIV infection and 1 patient with Crohn's disease was treated with corticosteroids; the remaining patients had no clinical evidence of depressed immune function. CONCLUSIONS: The incidence of vulvar ISCC in young women has increased over time; this increase cannot be accounted for by ISCC in immunocompromised patients. The overall disease outcome was excellent, with 2 of 21 patients dead of disease. Most tumors were associated with HPV, but cases of ISCC in the absence of HPV, and associated with differentiated VIN, were encountered. p53 staining of the basal layer can aid in recognition of differentiated VIN while MIB-1 staining within the upper layers of the squamous epithelium is consistently present in warty and basaloid VIN, but not in differentiated VIN.     

p53Mutations and Presence of HPV DNA Do Not Correlate with Radiosensitivity of Gynecological Cancer Cell Lines

Objective.The correlation betweenp53tumor suppressor gene mutations and the presence of high-risk human papillomavirus (HPV) DNA with thein vitroradiosensitivity of gynecological malignancies was studied in 26 cell lines derived from gynecological cancers of 23 patients.Methods.Comparison of the intrinsic radiosensitivity was performed with mean inactivation dose (D?) determined with the 96-well plate clonogenic assay.p53mutations were investigated with polymerase chain reaction and single-strand conformation polymorphism (PCR–SSCP) analysis and direct DNA sequencing, and the presence of HPV DNA was studied with PCR using HPV consensus primers.Results. p53mutations were found in 6 of 10 vulvar squamous cell carcinoma (SCC) lines. Nine vulvar and 1 vaginal SCC cell lines were HPV DNA negative and 1 vulvar cell line was HPV 16 positive. All 4 cervical SCC lines were HPV positive and possessed the wild-typep53.Three cell lines expressed HPV 16 and 1 HPV 68. Among 10 endometrial cancer cell lines, 2 cell lines with mutantp53and 1 HPV 16 positive cell line were found. No correlation could be demonstrated between inactivation of thep53gene and radiosensitivityin vitro;the cell lines were evaluated as one group or according to their anatomical origin or histology.Conclusion.Our results indicate that inactivation of thep53gene through mutation or binding with HPV DNA does not increase the resistance of gynecological malignancies to ionizing radiationin vitro.     

Mutation and expression of the TP53 gene in early stage epithelial ovarian carcinoma

OBJECTIVE: The early natural history of epithelial ovarian carcinoma remains poorly understood. Mutation of the TP53 gene is common in advanced-stage (III-IV) ovarian cancers, but less well described in early stage (I-II) tumors. The purpose of this study was to perform a comprehensive analysis of TP53 mutation and p53 expression status in early stage ovarian carcinomas. METHODS: Seventy-three cases of various histologic types, including 46 stage I and 27 stage II tumors, were subjected to direct sequence analysis of the entire TP53 coding region and exon-intron junctions as well as immunohistochemical assessment of p53 expression. RESULTS: Overall, mutations were identified in 24 of 73 (34%) cases. However, a significant difference in the distribution of mutations among histologic types was observed; TP53 mutations were present in 14 of 21 (67%) serous cancers and 11 of 52 (21%) non-serous cancers (P = 0.0002). Mutations were equally common between stage I and stage II tumors of serous histology. With respect to the correlation between TP53 mutation and p53 immunopositivity, the sensitivity (58%), specificity (71%), positive predictive value (64%), and negative predictive value (83%) were not sufficiently robust to justify use of p53 expression as a surrogate or screen for mutation. CONCLUSIONS: These data indicate that TP53 mutation is common in early stage ovarian carcinomas of serous histology, with a mutation frequency comparable to that reported for advanced-stage tumors, and is therefore likely to occur early in the progression of the most common histologic variant of ovarian carcinoma.     

Concordance of gross surgical and final fixed margins in vulvar intraepithelial neoplasia 3 and vulvar cancer

OBJECTIVE: To prospectively evaluate the concordance of initial surgical vulvar margins and final fixed margins and to determine the amount of microscopic pathology of grossly negative margins in women with vulvar intraepithelial neoplasia (VIN) 3 or vulvar carcinoma. STUDY DESIGN: Women with VIN 3 or vulvar carcinoma undergoing surgical excision were identified. Prior to excision, acetic acid was used to highlight the lesions, and 2 sutures were placed, 1 at the edge of gross disease and another 1 cm distal from the first. After specimen removal and fixation, the distance between sutures and microscopic involvement of VIN was determined. RESULTS: Twenty-seven women were enrolled; however, only 19 had final fixed specimens that could be accurately measured. The median fixed distance of the vulvar margin was 0.85 cm (mean, 0.83; SD, 0.19) as compared to the gross, 1-cm margin (p = 0.001). Three subjects (16%) had microscopic involvement by VIN 3 in the grossly negative epithelium between the 2 sutures, but none had a positive peripheral margin. CONCLUSION: The gross surgical margin after vulvar resection is reduced by 15% when measured in its final fixed state, and a grossly negative 1-cm margin will seldom harbor significant disease.     

p53 gene mutation is rare in human cervical carcinomas with positive HPV sequences

Chromosome 17p allelic losses and concurrent p53 mutations have been demonstrated in various human cancers. We therefore investigated the presence of chromosome 17p allelic loss and possible concurrent p53 mutation in 29 Korean cases of cervical carcinoma by restriction fragment length polymorphism (RFLP) analysis and polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) over the region from exon 4 to exon 9 of the p53 gene. We also examined the expression of p53 in paraffin tissues by immunohistochemical staining and determined the incidence of human papillomavirus (HPV) sequences in the same tissues by multitype PCR analysis to correlate them to the allelic loss on chromosome 17p13 and p53 mutation. In the analysis of 29 cases, loss of heterozygosity (LOH) was observed in eight (40%) cases out of 20 informative cases and p53 mutation was observed in only one case (3.4%) at exon 5. So in the majority of cases with LOH on 17p in this series, mutation of p53 gene appeared to be rare. But we obtained three cases (10.3%) of positive immunoreactivity from 29 cases. Those cases may carry mutations outside of the regions examined by PCR-SSCP. HPV DNA was detected in 27 of 29 cases (93.1%). HPV types 8, 11, 16, and 18 were detected in the samples we tested, while only two (7.4%) out of 27 HPV positive cases exhibited overexpression for p53 without any demonstrable p53 mutation upon PCR-SSCP. These results suggest that HPV infection may play a role in inactivating wild-type p53 protein in cervical carcinomas. In conclusion, mutation and overexpression of p53 gene appear to be rare, particularly in cases of cervical carcinoma associated with positive HPV sequence.     

Intraepithelial neoplasia and early stage vulvar cancer. Epidemiological, clinical and virological observations.

OBJECTIVE: Evaluation of vulvar intraepithelial neoplasia (VIN) and early vulvar cancer risk factor occurrence, frequency, localization and development. STUDY DESIGN: A clinical study carried out on 293 women aged 23-76 years with VIN and vulvar cancer stage I and in a control group of 115 cytologically and colposcopically negative women. METHODS: Clinical, colposcopic and morphological evaluation of the localization of VIN and vulvar carcinoma stage I concomitant with intraepithelial neoplasia in other parts of the lower genital tract. Anamnestic inquiry regarding risk factors. In situ hybridisation technique for HPV detection. Thomson's method for blood serum vitamin A level assessment. RESULTS AND CONCLUSION: An increased frequency of VIN and Ca stage I, especially in young women, has been observed in the past 15 years. In a group of young women under 45 years of age, those lesions were multifocal in 43 cases (63.2%), and unifocal in 25 patients (36.8%). In women over 45 years of age, multifocal lesions occurred in 35 (31.8%), and unifocal in 75 patients (68.2%). HPV infections concomitant with VIN and vulvar cancer stage I occurred in 61.5% of young women and in 17.5% of older females. VIN and stage I vulvar carcinoma coexisted with cervical intraepithelial neoplasia and cervical and/or vaginal cancer in 14 women (7.9%). The risk factor for VIN and early vulvar carcinoma occurrence in young women was different than in older patients. Long-term follow-up of VIN 1 and VIN 2 showed that in over 1/3 of cases the lesions were persistent or recurred after a transient remission. Progression depended not only on dysplasia stage, but also on histological pattern.     

Effect of human papillomavirus vaccines on vulvar, vaginal, and anal intraepithelial lesions and vulvar cancer

OBJECTIVE: Human papillomavirus (HPV) is a necessary cause for cervical cancer, and it has been associated with vulvar and vaginal cancer and vulvar (VIN) and vaginal (VaIN) and anal (AIN) intraepithelial neoplasia. We assessed the prevalence of HPV (and the types) to estimate the possible effect of a HPV vaccine on lower genital tract disease prevention. METHODS: Two hundred fifty-eight samples of VIN, VaIN, AIN, and vulvar cancer from 241 women were included in the study. The diagnosis of surgical samples was made using published histomorphologic criteria. The DNA was extracted for HPV detection and typed using polymerase chain reaction and sequencing. RESULTS: The analyses were performed on 210 intraepithelial neoplasia samples (VIN2/3, VaIN2/3, AIN2/3) and 48 vulvar carcinoma samples. Human papillomavirus DNA was detected in 92%, 91%, 89%, and 60% of the VIN, VaIN, AIN, and vulvar carcinoma samples, respectively. High-risk HPV types 16 or 18 were detected in 76%, 64%, 81%, and 42% of the VIN2/3, VaIN2/3, AIN, and vulvar carcinoma samples. Women with HPV-positive samples were younger than those with HPV-negative samples (46 years compared with 55 years and 51 years compared with 61 years, for the VIN2/3 and vulvar carcinoma samples, respectively). Human papillomavirus-positive vulvar carcinoma was more frequent in women aged younger than 56 years (77%), than in those aged 56 years or older (41%). CONCLUSION: Based on the data obtained in this study, widely-implemented prophylactic HPV vaccination could make an important contribution to the reduction of the risk for cervical cancer and could also prevent about half the vulvar carcinomas in younger women and about two thirds of the intraepithelial lesions in the lower genital tract. LEVEL OF EVIDENCE: II-3.