人乳头状瘤病毒(HPV)和P53蛋白在尖锐湿疣与阴茎癌中的表达

目的： 探讨ＨＰＶ 及Ｐ５３蛋白在尖锐湿疣和阴茎鳞癌中的表达情况。　方法： 在常规病理确定诊断后,用ＳＰ免疫组化法显示标本内的ＨＰＶ 抗原和Ｐ５３ 蛋白。　结果：尖锐湿疣２１ 例,ＨＰＶ标记均阳性,Ｐ５３ 蛋白标记５例见低度表达。阴茎鳞癌１９ 例,ＨＰＶ 标记阳性２例,Ｐ５３ 蛋白标记１２例见过度表达（阳性率６３％ ）,其中６ 例强阳性。　结论：尖锐湿疣与ＨＰＶ 感染有关,少数见Ｐ５３ 蛋白低度表达;而阴茎鳞癌中常见Ｐ５３蛋白过度表达,少数可能与ＨＰＶ感染有关。     

Human Papilloma Virus,Histopathological, and Molecular Subtyping in Penile Cancer: Relevance for Prognosis and Therapy

ContextBased on the new 2016 World Health Organization classification, penile carcinomas are subdivided into several histopathological subtypes which are associated with human papilloma virus (HPV) infection status.ObjectiveThe primary objective of this review is to describe the subtyping of penile carcinomas according to the new World Health Organization classification based on HPV infection status and histopathological features and to correlate this to patient outcome. In addition, the current knowledge about the role of HPV infection in tumorigenesis as well as relevant molecular alterations will be presented.Evidence acquisitionA systematic literature search was conducted using PubMed to identify original articles, review articles regarding penile cancer/carcinoma, HPV, histopathological subtypes, prognosis, and molecular alterations. Articles published between 1989 and 2017 were reviewed and selected with the consensus of all the authors.Evidence synthesisPenile squamous cell carcinomas (SCC) are the most common penile tumours. They are divided into HPV-related and HPV-unrelated subtypes based on the frequency of high-risk HPV infection. Histopathological subtype but not the HPV-status alone is associated with clinical outcome. High-risk HPV infection occurs in about 50% of penile SCC and varies worldwide. Although some specific molecular alterations including copy number alterations or mutations (most frequently CDKN2A and p53) and changes in signalling pathways could be identified, molecular pathogenesis considering HPV infection is not fully understand.ConclusionsThe histopathological subtype should be documented and considered for prognostic evaluation in patients with penile SCC. The prognostic value of HPV infection has to be investigated in larger cohorts considering the histopathological subtype. Further molecular studies have to identify relevant molecular subtypes and to develop type-specific targeted therapies.     

人乳头瘤病毒DNA相关序列与p53基因突变对大肠癌生物学行为的影响50例报告

目的探讨人乳头瘤病毒（ＨＰＶ）ＤＮＡ相关序列与ｐ５３基因突变及ｐ５３蛋白表达的关系及其对大肠癌生物学行为的影响。方法采用ＰＣＲ方法检测大肠癌及癌旁组织、肝转移灶中ＨＰＶＤＮＡ相关序列。并应用ＰＣＲＳＳＣＰ及免疫组化技术分别检测ｐ５３基因突变及ｐ５３蛋白表达。结果在５０例大肠癌中,检出ＨＰＶ１６、１８ＤＮＡ相关序列２６例（５２０％）,其中ＨＰＶ１６ＤＮＡ４例（８０％）,ＨＰＶ１８ＤＮＡ２２例（４４０％）。ｐ５３基因突变率为５６０％。ｐ５３蛋白表达阳性率为４２０％。ＨＰＶＤＮＡ相关序列与ｐ５３基因突变及ｐ５３蛋白表达呈正比关系。结论ＨＰＶＤＮＡ相关序列可促进细胞转化、致ｐ５３基因突变、抑制细胞的凋亡,与大肠癌的发生发展有密切关系。     

The expression of key cell cycle markers and presence of human papillomavirus in squamous cell carcinoma of the tonsil

BACKGROUND: Chemical carcinogens induce squamous cell carcinoma (SCC) of the head and neck by targeting the p53 and the retinoblastoma (pRb) pathways. Human papillomavirus (HPV) might have an etiologic role in these cancers at particular sites. Few studies have compared cell cycle protein expression in HPV-positive and HPV-negative tumors in this region. METHODS: Fifty tonsil SCCs were analyzed for HPV by PCR and for expression of cell cycle proteins (p53, pRb, p16(INK4A), p21(CIP1/WAF1), p27(KIP1), and cyclinD1) by immunohistochemistry. RESULTS: HPV was present in 42%; almost all were type 16. There were statistical associations between HPV positivity and reduced expression of pRb and cyclinD1, overexpression of p16, and younger patient age. Tumor with down-regulated p27 tended to have down-regulated pRb and p21. CONCLUSIONS: HPV-positive tonsil SCCs have distinct molecular pathways. Their association with younger patient age suggests that they are biologically distinct from HPV-negative tumors.     

Correlation between p53 mutations and HPV in bilharzial bladder cancer

Alterations of the p53 tumor suppressor gene are the most common genetic changes detected in human cancers as well as in papillary and invasive bladder cancer. Several studies have demonstrated an association between HPV infection and urological malignancies. In the present work, the p53 gene status was studied together with the frequency of HPV in 99 cases of Bilharzial bladder cancer [BBC] in Egypt and both were correlated to the clinicopathological features of the patients. SSCP and sequencing were used to screen the p53 gene for mutations at exons 4-10 and IHC was performed to detect protein overexpression. PCR was used for detection and typing of HPV-DNA in tumor samples. p53 mutations were detected in 33.3% of the studied cases whereas protein overexpression was detected in 35.6% of the cases. The highest concordance rate was observed in cases harboring mutations at exon 4 [87.5%]. Bilharzial infestation was obvious in 72.2% of the cases that showed mutations. Exon 8 showed the highest rate of mutation [32%] followed by exons 4 and 5 [22% each]. The commonest mutational event was G:C transversion [15/50] especially at CpG dinucleotides. A mutational hot spot was detected at exon 4, codons 72-73. HPV-DNA was detected in 48.97% of the cases the majority of which [64.6%] were of type 16. Significant correlation was found between p53 mutation and the pathological stage as well as p53 overexpression and tumor grade. Our results demonstrate that the mutational spectrum in BBC is different from that of bladder cancer in Western countries in many aspects and suggest an etiological role of HPV in this type of neoplasm. However, both HPV infection and p53 gene abnormalities may contribute to Bilharzial bladder carcinogenesis in an independent way.     

Downregulation of miR-145 is associated with perineural invasion in penile carcinoma

BackgroundHuman papillomavirus (HPV) infection is a risk factor for penile cancer (PC). The miR-145 expression has been correlated to this virus genomic amplification. In this context, this work aims to determine the expression level of miR-145 in penile tumors infected by high-risk HPV and correlate it with the clinicopathological characteristics of the tumor and protein expression of p53.MethodsFormalin-fixed paraffin-embedded from 52 patients with PC, at diagnosis and prior to any cancer treatment, were obtained. HPV identification was performed by nested type PCR, and miR-145 expression was obtained by qRT-PCR. Immunohistochemical analysis of p53 and Ki-67 was performed.ResultsTumoral miR-145 expression was significantly lower compared to adjacent tissue. Additionally, there was a significant reduction of miR-145 expression in invasion perineural, histological associated HPV, and absence of p53 expression in positive HPV cases. HPV infection was detected in 86.5%, the most frequent HPV16. Reduced disease-free survival was observed in patients with low expression of miR-145.ConclusionsOur data suggest that the underexpression of miR-145 may be triggered by HPV action, decreasing protein expression of p53, and being correlated with perineural invasion. Therefore, the deregulation of miR-145 provides clues as to the potential role in penile carcinogenesis and is also a potential candidate for validation in noninvasive samples.     

乳腺癌患者高危型人乳头状瘤病毒感染、p53蛋白表达及淋巴结转移分析

目的分析乳腺癌患者高危型人乳头状瘤病毒（HPV）感染、p53蛋白表达和淋巴结转移率。 方法选取2016年5月至2018年5月于西北妇女儿童医院治疗的乳腺癌患者共90例作为观察组,选取同期于本院治疗乳腺增生患者90例作为对照组。对癌组织行HPV基因分型和p53蛋白检测,增生组织行HPV基因分型。观察入组患者高危型HPV感染率,分析乳腺癌患者肿瘤大小、TNM分期、淋巴结转移率与p53蛋白表达等。 结果观察组患者中共60例发生高危型HPV感染,对照组中共36例发生高危型HPV感染,观察组患者中HPV16、18基因型感染率分别为21.11%（19/90）和22.22%（20/90）,均高于对照组[2.22%（2/90）和2.22%（2/90）],差异均有统计学意义（χ² = 7.108、P = 0.001,χ² = 8.063、P = 0.001）。观察组HPV阳性患者淋巴结转移率为93.33%（56/60）,显著高于HPV阴性患者（21/30、70.00%）,差异有统计学意义（χ² = 4.072、P = 0.002）。观察组患者中p53蛋白阳性者39例（39/90、43.33%）,其中肿瘤大小≤ 2 cm者27例（27/39、69.23%）、TNM分期Ⅰ期、Ⅱ期、Ⅲ期和Ⅳ期分别占15.38%（6/39）、30.77%（12/39）、35.90%（14/39）和17.95%（7/39）,淋巴结转移率为82.05%（32/39）;p53蛋白阴性患者51例（51/90,56.67%）,肿瘤大小≤ 2 cm者35例（35/51、68.63%）、TNM分期Ⅰ期、Ⅱ期、Ⅲ期和Ⅳ期分别占7.84%（4/51）、19.61%（10/51）、49.02%（25/51）和23.53%（12/51）,淋巴结转移率为86.27%（44/51）,p53蛋白阴性组与p53蛋白阳性组患者肿瘤大小、TNM分期及淋巴结转移率差异均无统计学意义（χ² = 0.682、P = 0.462,χ² = 0.491、P = 0.507,χ² = 0.572、P = 0.461）。 结论检测乳腺癌患者高危型人乳头状瘤病毒感染、p53蛋白表达及淋巴结转移率有助于对乳腺癌的诊疗;p53蛋白阳性率下降可能促进HPV感染相关肿瘤的发生和发展。     

Expression of p16(INK4A), p53, and Rb proteins are independent from the presence of human papillomavirus genes in oral squamous cell carcinoma.

BACKGROUND: The overexpression of p16(INK4A) and suppression of p53 and Rb proteins are key features of oncogenic transformation by human papillomaviruses (HPV) in anogenital cancers. HPV genomes are often detected in cancers of the oral cavity, but it is unclear whether HPV has a specific oncogenic role there. OBJECTIVES: The objectives of the study were to investigate the expression of p53, Rb, and p16(INK4A) proteins and identify HPV infection and viral integration into the host genome. METHODS: Seventy-nine cases of oral squamous cell carcinoma (OSCC) were studied by immunohistochemistry. Polymerase chain reaction (PCR) was performed to identify HPV DNA from the samples. The results were correlated with clinical data. RESULTS: Thirty-three cases were HPV positive for high-risk HPV (HR-HPV) types, of which 27 harbored HPV16. In 25 of 27 HPV16-positive tumors, the HPV16 genome was fully integrated into the host genome, as evidenced by the lack of PCR-amplifiable E2 gene sequences. Forty-five patients were p53 overexpressing, 20 with HR-HPV-positive and 25 with HR-HPV-negative tumors. p16(INK4A) protein was overexpressed in 4 of 31 HR-HPV-positive and 9of 45 HR-HPV-negative cases. Twenty-six of 32 HR-HPV-positive and 37 of 44 HR-HPV-negative samples exhibited pRb nuclear staining. These differences between HR-HPV-positive and -negative tumors were not statistically significant. No correlation was found between these biological factors and tumor location, stage, differentiation grade, or alcohol or tobacco abuse. CONCLUSIONS: A tumor immunophenotype, similar to HPV-related anogenital cancers, is not present in OSCC and highly oncogenic HPV types are therefore unlikely to be specific or independent risk factors for oral cancer.     

Presence of human papillomavirus DNA and abnormal p53 protein accumulation in lung carcinoma.

BACKGROUND: In some carcinomas inactivation of the tumour suppressor gene product p53, either by point mutation or indirectly by the human papillomavirus (HPV), has been suggested as two alternative routes to malignant transformation. To test this hypothesis in lung tumours, 43 lung carcinomas were analysed by in situ hybridisation and polymerase chain reaction (PCR) for the presence of HPV DNA, and the results were compared with p53 protein immunohistochemical analysis. METHODS: The presence of HPV DNA in lung carcinoma was detected by nucleic acid in situ hybridisation for HPV types 6, 11, 16, 18, 31, and 33 using nonradioactively labelled DNA probes. Polymerase chain reaction (PCR) analysis was performed on all cases showing positive HPV DNA labelling by in situ hybridisation and in an additional 13 negative cases. Abnormal nuclear accumulation of the p53 protein was revealed by immunohistochemistry using the avidin-biotin-peroxidase complex method and a CM-1 polyclonal anti-human p53 antibody and a monoclonal mutation-specific Pab 240 p53 antibody. RESULTS: HPV DNA was found by in situ hybridisation in 13 lung carcinomas (30%). In all these cases subtype-specific HPV DNA could also be detected by PCR. Abnormal p53 protein accumulation was seen in 21 of the 43 carcinomas (49%), of which 18 were HPV negative. Twelve (57%) of the CM-1 positive cases were also positive for the mutation-specific antibody Pab 240. There was an obvious inverse relationship between the presence of papilloma viral DNA and abnormal p53 protein accumulation. CONCLUSIONS: p53 plays an important part in the development of lung carcinomas and, in some cases, HPV may contribute to it by binding and inactivating the p53 protein.     

Human Papillomavirus and Overexpression of P16INK4a in Nonmelanoma Skin Cancer

BACKGROUND: P16INK4a overexpression has been identified as a specific biomarker in high-risk human papillomavirus (HPV)-infected cervical (pre)cancer lesions. OBJECTIVE: To evaluate the overexpression of this cyclin-dependent kinase inhibitor in skin tumors depending on HPV infections, we analyzed normal skin, benign skin disease, and skin cancer specimens. METHODS: Biopsies of 23 patients with normal histology (3), psoriasis (2), verrucae vulgaris (2), actinic keratoses (5), squamous cell carcinoma (SCC) in situ (3), Bowen's carcinoma (1), and SCC (7) were analyzed. Specimens of 23 patients were immunostained using the monoclonal antibody E6H4 specific for p16INK4a. HPV status was assessed by a polymerase chain reaction (PCR) system to detect all currently known HPV types. MY (MY09/MY11 and MYN9/MYN10)-, CP (CP65/CP70 and CP66/CP69)-nested PCR, and three single PCR methods CN1, CN3, and CN4 were used in a first step, and HPV typing was performed by restriction fragment length polymorphism analysis. Only beta-globin-positive patients were included in this study. RESULTS: HPV DNA was detected in all actinic keratoses, SCC in situ, Bowen's carcinoma, and SCC, in 50% (one of two) of verrucae vulgaris, in 66% (two of three) of normal skin, and in none of two psoriasis. P16INK4a expression was not detected in normal skin, psoriasis, and verrucae vulgares. Overexpression of p16INK4a was detected in a subset of dysplastic cells (10% to 80%) of all skin (pre)cancer lesions such as actinic keratoses, SCC in situ, Bowen's carcinoma, and SCC infected with HPV independent of sun exposure. CONCLUSION: P16INK4a appears to be overexpressed in a portion of dysplastic cells from actinic keratoses and SCC. Further studies to examine the association of HPV infection and the overexpression of p16INK4a are warranted.     

Detection of human papillomavirus DNA and expression of p16, Rb, and p53 proteins in small cell carcinomas of the uterine cervix

Human papillomavirus (HPV) has been implicated as an etiologic agent for the development of primary small cell carcinoma of the uterine cervix, a rare but highly aggressive malignancy. It has been shown that the HPV E6 and E7 oncoproteins are able to inactivate the tumor suppressor functions of p53 and Rb. In squamous cell carcinoma and adenocarcinoma of the cervix, HPV infection is also associated with overexpression of p16, a cyclin-dependent kinase inhibitor. In this study, 22 cases of primary small cell carcinoma of the uterine cervix were subjected to broad-spectrum HPV DNA amplification and typing, and immunohistochemically examined for the expression of p16, Rb, and p53 proteins. The results show that HPV DNA was detected in every case (100%), with 18 cases (82%) harboring type 18. The tumor cells exhibited strong nuclear staining for p16 in 20 cases (91%). This was associated with a complete loss of Rb nuclear staining in tumor cells in 16 cases (73%). The p53 protein was essentially undetectable in all cases. In contrast, HPV DNA was not detected in 9 colorectal and 8 urinary bladder small cell carcinomas included in this study for comparison. While similar p16 and Rb expression patterns were observed in these HPV-negative tumors, a different expression pattern for p53 was noted where strong nuclear staining was seen in 8 cases (47%; P = 0.0004 compared with cervical tumors). These observations indicate that different mechanisms are involved in the pathogenesis of small cell carcinomas of the uterine cervix and support the notion that nuclear p16 overexpression serves as an indication of Rb defunctioning in tumor cells, which may or may not result from high-risk HPV infection.     

Human papillomaviruses associated with cervical intraepithelial neoplasia. Great diversity and distinct distribution in low- and high-grade lesions.

All together, 30 genital human papillomavirus (HPV) types have been characterized so far. To evaluate the importance of HPV diversity in associated cervical diseases, we analyzed 188 biopsy specimens obtained from patients with a recent diagnosis of cervical HPV infection or intraepithelial neoplasia (CIN). Of these 188 specimens, 116 were classified as low-grade CIN (48 cases), high-grade CIN (53 cases), condylomata acuminata (10 cases), flat condylomas (five cases). Seventy-two specimens were considered nondiagnostic. Using probes specific for 18 genital HPV types, HPV DNA sequences were detected by Southern blot hybridization in 100 lesions and 21 nondiagnostic specimens. When further analyzed by the polymerase chain reaction, eight HPV-negative biopsy specimens, four CIN, and four nondiagnostic specimens were positive. Of the 129 positive biopsy specimens, 92 contained at least one of 18 known HPV types and 37 HPV that have not yet been identified. Nine specimens had more than one type. Thirteen HPV types were identified in CIN. The detection rate of HPV 16 increased from 21% in low-grade CIN to 57% in high-grade CIN. HPV 18 was detected in only 3% of CIN; HPV 31, 33, and 35 were found in 8%. HPV 30, 39, 45, 51, 52, 56, 58, and 61 were detected in 44% of low-grade CIN but in only 8% of high-grade CIN. Unidentified HPV were detected in about 25% of low-grade and high-grade CIN. Fifty-seven CIN positive for at least one HPV type were further analyzed by in situ hybridization. Thirty-five (65%) biopsy specimens were positive, including 21 of 24 low-grade CIN and 14 of 33 high-grade CIN. Ten of the 13 previously identified HPV types were detected. Thus, CIN represents an heterogeneous disease from a virologic viewpoint. This fact could explain their variable clinical evolution.     

Combining routine morphology,p16INK4a immunohistochemistry,and in situ hybridization for the detection of human papillomavirus infection in penile carcinomas: A tissue microarray study using classifier performance analyses

ObjectivesInfection by high-risk human papillomavirus (HR-HPV) plays an important role in the pathogenesis of penile cancer in approximately 50% of the patients. The gold standard for human papillomavirus (HPV) detection is the polymerase chain reaction (PCR) assay. However, technical requirements and associated costs preclude the worldwide use of PCR assays on a routine basis. Herein, we evaluated the predictive abilities of tumor morphology, immunohistochemistry for p16^INK4a expression, and in situ hybridization (ISH) for HR-HPV detection in defining HPV status, as established by PCR.Materials and methodsTissue samples from 48 patients with HPV-positive penile squamous cell carcinoma (SCC) were included in 4 tissue microarrays (TMA).ResultsSensitivities and specificities were as follows: tumor morphology, 70% and 68%; p16^INK4a immunohistochemistry, 65% and 90%; HR-HPV ISH, 47% and 100%. Regarding combinations of the predictors, the best performance was seen when HR-HPV ISH and p16^INK4a immunohistochemistry were combined, regardless of the tumor morphology: sensitivity, 88%; specificity, 64%; area under the receiver-operating characteristic (AUC) curve, 0.83. Combinations of tumor morphology with p16^INK4a immunohistochemistry or with HR-HPV ISH performed similarly well.ConclusionsIn penile SCC, both p16^INK4a immunohistochemistry and ISH for HR-HPV increase the predictive ability of routine morphology in defining HPV status. These tests can be interpreted differentially, depending on the necessity of a higher sensitivity or a higher specificity. For research/screening studies, we recommend combining tumor morphology, p16^INK4a immunohistochemistry, and HR-HPV ISH. To increase sensitivity, positivity in any of these predictors should be considered as indicative of HPV infection. For routine diagnosis of clinical cases, criteria should be more stringent, and, to achieve the highest specificity in classifying a case as HPV-related, all predictors should be consistently positive. The data generated in the present study could be used in algorithms for defining HPV status in penile carcinomas.     

The expression of proliferating cell nuclear antigen,p53, p21, and apoptosis in primary gastric lymphoma

BACKGROUND: We have investigated the correlations among the expression of proliferating cell nuclear antigen (PCNA), p53, p21, and the prognosis of primary gastric lymphoma, and we have investigated apoptosis by using transferase-mediated deoxyuridine triphosphate nick-end labeling staining. METHODS: A retrospective study was performed on 33 cases of primary gastric lymphoma that were surgically resected. Histopathologic examination was undertaken according to the Working Formulation classification. Immunohistochemical staining was performed by using the avidin-biotin-peroxidase complex method, with anti-PCNA antibody, anti-p53 antibody, and anti-p21 antibody. Apoptosis was quantified in situ from paraffin-embedded specimens by using an in situ apoptosis detection kit. RESULTS: On histologic grade, the survival rate of high-grade type was significantly lower than that of low-grade type and than that of intermediate-grade type. The PCNA index of low-grade type was significantly lower than that of intermediate-grade type and than that of high-grade type. Those patients who experienced recurrence were both p53(+) and p21(-). Two of these patients were receiving postoperative chemotherapy but nevertheless experienced recurrence at 18 months and at 5 months after surgery. The PCNA index of p53(+) p21(-) cases was significantly higher than that of p53(-) p21(+) cases. The apoptotic count of p53(-) p21(+) cases was significantly higher than that of p53(+) p21(-) cases. CONCLUSIONS: The Working Formulation classification and the PCNA index were each significant factors predicting the prognosis of primary gastric lymphoma. The prognosis of p53(+) p21(-) cases was poor, and the apoptotic count of p53(+) p21(-) cases was low. Accordingly, the effect of combined chemotherapy for those cases was thought to be poor.     

p53 as a new prognostic factor for lymph node metastasis in penile carcinoma: analysis of 82 patients treated with amputation and bilateral lymphadenectomy

PURPOSE: Gold standard treatment for invasive penile carcinoma remains amputation and lymphadenectomy. This procedure has high morbidity and new prognostic factors on the incidence of metastasis would help select candidates to lymphadenectomy. Mutations in the p53 gene common in several neoplasms can be related to the prognosis. We studied 82 patients with penile carcinoma staged according to the 1978 TNM system who underwent amputation and bilateral lymphadenectomy to evaluate the prognostic value of immunohistochemical p53 staining in the primary tumor. MATERIALS AND METHODS: Immunoreactivity of p53 was studied with other clinicopathological variables, including patient age, stage, histological grade, tumor thickness, lymphatic and venous embolization, corpora cavernosa, corpus spongiosum and urethral infiltration, and human papillomavirus (HPV) status. We also determined its association with lymph node metastasis, the survival rate and the risk of death. In addition, we studied the association of p53 and HPV DNA with prognosis. All slides were reviewed by 1 pathologist. HPV DNA was detected by polymerase chain reaction using GP5/6+ generic primers. p53 expression was measured by immunohistochemical testing with monoclonal Clone DO-7 mouse anti-human p53 protein antibody (Dako A/S, Glostrup, Denmark). The Cox regression hazards method was used for multifactorial analysis. RESULTS: Nuclear accumulation of p53 was detected in 34 of 82 samples (41.5%). Clinical lymph node N stage (p = 0.045), lymphatic (p <0.001) and venous (p = 0.04) embolization by neoplastic cells, p53 positivity (p = 0.012) and p53 grade (p = 0.004) were significantly associated with lymph node metastasis. Followup was 0.1 to 453 months (mean 88.7). Multivariate analysis revealed that only lymphatic embolization (relative risk 9.4, 95% confidence interval [CI] 2.8 to 31.6) and p53 positivity (relative risk 4.8, 95% CI 1.6 to 14.9) were independent factors for lymph node metastasis. Patients with negative p53 had significantly better 5 and 10-year overall survival than those in whom tumors stained positive for p53 (64.5% and 54.6% versus 30.2% and 26.4%, respectively, p = 0.009). When tumors were p53 positive and HPV DNA positive, overall survival was worse. Multivariate analysis revealed that only age (relative risk 2.9, 95% CI 1.6 to 5.1) and lymph node metastasis (relative risk 3.2, 95% CI 1.8 to 5.8) were independent risk factors for death. CONCLUSIONS: Immunoreactivity of p53 is an independent factor for lymph node metastasis. The association of positive p53 with positive HPV DNA was related to a worse prognosis.     

Role of human papillomavirus in squamous cell metaplasia-dysplasia-carcinoma of the rectum

Primary colorectal squamous cell carcinoma (SCC) and squamous dysplasia are uncommon and little is known about their pathogenesis. Most have been reported in association with ulcerative colitis and other chronic disease states. Although cervical and anal SCC have been strongly linked to human papillomavirus (HPV) infection, the role of HPV in rectal squamous carcinoma has not been well-examined. We evaluated 3 cases of primary rectal SCC for the presence of high-risk HPV by immunohistochemistry for p16(INK4A), in situ hybridization, and polymerase chain reaction. HPV type 16 was detected by polymerase chain reaction in all cases. In addition, all cases exhibited diffuse strong reactivity for p16(INK4A) and punctate nuclear staining by Ventana HPVIII in situ hybridization. The presence of HPV 16 in all three cases suggests that high-risk HPV infection is a risk factor for rectal SCC, particularly in patients with underlying chronic inflammatory disease processes or altered immune status. Further studies are warranted to determine if SCC occurring more proximal in the colon are also HPV-dependent or occur via another, HPV-independent pathway.     

人骨肉瘤中p53、p21~(WAF1/CIP1)和增殖细胞核抗原的表达及相互关系

目的 探讨细胞周期相关基因p21~(WAF1/CIP1)、p53和增殖细胞核抗原(PCNA)在人骨肉瘤中的表达和相互关系.方法 应用LSAB法,对40例骨肉瘤、20例骨软骨瘤患者的病变组织和20例正常人骨组织分别以p53、p21~(WAF1/CIP1)、PCNA蛋白的单克隆抗体作免疫组织化学检测.结果 骨肉瘤组织中p53、p21~(WAF1/CIP1)、PCNA蛋白的阳性反应率和阳性反应强度均明显高于骨软骨瘤和正常骨组织,其差异有统计学意义(P<0.05).p53标记指数与p21~(WAF1/CIP1)标记标数以及p21~(WAF1/CIP1)与PCNA标记指数间呈正相关(t值分别为2.93和4.20,P<0.01).结论 骨肉瘤组织中有p53、p21p21~(WAF1/CIP1)和PCNA的过表达,其表达的强度与肿瘤的恶性程度和预后相关.     

Human papilloma virus DNA and p53 mutation analysis on bladder washes in relation to clinical outcome of bladder cancer

OBJECTIVES: High-risk human papilloma virus (HPV) types stimulate degradation and deactivation of protein associated with the p53 tumour suppressor gene via the ubiquitin-dependent pathway. For a long time, changes of the p53 tumour suppressor gene have been correlated with poor clinical outcome in patients with superficial bladder cancer. We aimed to study the association between presence of (high-risk) HPV DNA, p53 status, and clinical outcome in bladder cancer patients. This study must be seen as a preliminary study to investigate this potentially important problem. MATERIAL AND METHODS: From 107 patients, 166 bladder wash samples were obtained. p53 status was determined by mutation analysis, HPV detection, and genotyping by the SPF(10)-LiPA assay. Clinical data were abstracted from the medical files. RESULTS: The prevalence of all-type and high-risk HPV infection in malignancies of the bladder was 15.2% and 8.1%, respectively. In high-grade tumours this prevalence was 18.2% and 10.6%, respectively. In grade 1, 2 and 3 tumours the infection rate of high-risk HPV types was 0%, 3.3%, and 10.6%, respectively (trend test: p=0.221). In Ta, T1, and T2-T4 tumours the high-risk HPV infection rate was 0%, 12.5% and 18.2%, respectively (trend test: p=0.045). In the p53 wild-type patients who showed progression, 1 of 9 patients had a high-risk type HPV infection. In the group of wild-type patients who showed no progression, 4 of 37 patients had a high-risk type HPV infection (odds ratio: 1.03; 95% confidence interval, 0.1-10.5). CONCLUSIONS: The data of this pilot study show the suggestion of a positive trend in the correlation between tumour grade/stage and high-risk type HPV infection. However, no additional risk for progression is found for p53 wild-type patients with a high-risk HPV infection.     

Prevalence of high-risk human papillomavirus DNA in nonkeratinizing (cylindrical cell) carcinoma of the sinonasal tract: a distinct clinicopathologic and molecular disease entity

Carcinomas of the nose and paranasal sinuses are a heterogeneous group of neoplasms that differ histologically, biologically, and clinically. Some of these tumors are known to harbor high-risk human papillomavirus (HPV) DNA. In an attempt to identify specific phenotypes associated with HPV infection, 39 cases of sinonasal carcinomas were evaluated by PCR for the presence of HPV DNA. The tumors were also studied with a panel of immunohistochemical stains, including p16, p53, and Ki-67 antibodies. Twenty-one cases were identified as keratinizing squamous cell carcinoma (KSCC) with a male-to-female ratio of 3:1. Eight cases were nonkeratinizing (cylindrical cell) carcinoma (NKCa) with a male-to-female ratio of 1:1. Ten cases were sinonasal undifferentiated carcinoma (SNUC), and 9 of these patients were men. HPV DNA, particularly type 16, was detected in 9 cases: 4 of 21 (19%) of KSCC, 4 of 8 (50%) of NKCa, and 1 of 10 (10%) of SNUC. In addition to a higher prevalence of HPV DNA in NKCa, the tumors also showed a distinct immunophenotype characterized by strong and diffuse staining for p16, high labeling scores for Ki-67, and negative or low reactivity to p53. On the other hand, KSCC and SNUC were either negative or weakly reactive to p16 antibodies. KSCC cases were more likely to be positive and more strongly reactive to p53 stain. Unlike KSCC, SNUC had high Ki-67 labeling scores. These observations suggest that NKCa of the sinonasal tract is a distinct histopathologic and molecular disease entity, which should be added to the list of upper aerodigestive tract tumors with strong etiologic relationship to high risk HPV.