Small-cell neuroendocrine carcinoma of the cervix. A human papillomavirus type 18-associated cancer.

Small-cell undifferentiated carcinomas comprise a rare but aggressive subset of uterine cervical neoplasms. Analogous to small-cell anaplastic carcinoma of the lung, these tumors frequently exhibit neuroendocrine differentiation. Although human papillomaviruses (HPV) types 16 and 18 are strongly associated with the development of cervical squamous carcinoma, there is as yet little information describing the relationship of these viruses to small-cell carcinomas. To address this question, we analyzed 20 cases of small-cell carcinoma of the cervix using in situ hybridization to detect HPV gene expression. In addition, immunohistochemistry was used to evaluate three markers of neuroendocrine differentiation. Eighteen of 20 tumors (90%) demonstrated some evidence of neuroendocrine differentiation; 17 of 20 (85%) expressed HPV type 16 or 18 messenger RNA. Of the neuroendocrine-positive cases, 14 of 18 expressed HPV 18 messenger RNA. In contrast, both of the cases with squamous differentiation were HPV 16 positive. These findings broaden the spectrum of HPV-associated cervical neoplasia and strongly suggest that HPV 18 is a viral type specifically associated with cervical small-cell neuroendocrine carcinomas.     

Detection of human papillomavirus in formalin-fixed, invasive squamous carcinomas using the polymerase chain reaction

We analyzed 88 formalin-fixed, paraffin-embedded invasive squamous carcinomas for human papillomavirus-related DNA sequences (HPV types 16 and 18) following in vitro gene amplification using the polymerase chain reaction. HPV DNA sequences were found in 35 of 50 (70%) carcinomas of the anogenital region, including four of four (100%) anal, six of eight (75%) vulvar, nine of 14 (64%) vaginal, two of five (40%) penile, and 14 of 19 (74%) cervical tumors. Nine of 25 (36%) oropharyngeal squamous carcinomas contained HPV DNA sequences, including four of 10 (40%) laryngeal, three of eight (38%) buccal, and two of seven (29%) glossal tumors. HPV DNA sequences were not found in 13 esophageal carcinomas. Of the 44 cases that contained viral DNA, HPV-16 was detected in 41 cases (93%) and HPV-18 in five cases (11%), while both types were found in two cases (one anal and one vulvar). HPV DNA sequences were found in 43 of 83 (52%) nonverrucous and in one of five (20%) verrucous carcinomas, but this difference was not significant. These findings demonstrate that HPV DNA sequences are more frequently associated with anogenital than oropharyngeal squamous carcinomas and can be readily detected in routinely processed tissues using the polymerase chain reaction.     

Detection of human papillomavirus in small cell carcinomas of the anus and rectum

Small cell carcinomas represent <1% of colorectal/anal carcinomas and have a poor prognosis. Anorectal squamous cell carcinomas are often associated with high-risk human papillomavirus (HPV) infection, similar to squamous and small cell carcinomas of the uterine cervix. In HPV infection, the oncoprotein E7 inactivates the tumor suppressor Rb, leading to p16 upregulation; however, in small cell carcinomas, the Rb pathway is often blocked by other mechanisms; thus, increased p16 may not indicate HPV infection. P16 immunohistochemistry (IHC) may have a limited role in evaluating small cell carcinomas for HPV infection. Formalin-fixed, paraffin-embedded tissue sections of previously diagnosed small cell carcinomas of the anus (n=5) and rectum (n=11) were subjected to IHC for p16, CDX2, and p63, followed by in situ hybridization for high-risk HPV. All (100%) cases of anal and rectal small cell carcinomas were positive for p16, and 100% of anal and 82% of rectal small cell carcinomas were positive for high-risk HPV. The majority of cases showed low or very low HPV copy numbers. In 6 cases, HPV was detected only by using the HPV-16 genotype-specific assay detecting very low copy numbers (1 to 2 viral copies). The majority of tumors expressed p63, which was more pronounced in the anal tumors. CDX2 expression was observed predominantly in rectal tumors. High-risk HPV can be detected using in situ hybridization in the majority of anorectal small cell carcinomas, which are uniformly p16 positive by IHC. HPV-targeted therapy could result in better control of these aggressive neoplasms.     

Human papillomavirus-associated early vulvar neoplasia investigated by in situ hybridization

Of 21 consecutive cases of early vulvar neoplasia studied at the Istituto Nazionale Tumori of Milan, 62% appeared to be related to papillomavirus infection. This conclusion is the result of the present study by in situ hybridization with DNA probes of human papillomavirus (HPV) 6/11, 16, and 18 and of previous ultrastructural and immunohistochemical investigations. The proportion of cases associated with HPV was 78.5% for those (11/14) with histologic evidence of viral infection and 33% for those without (2/6). HPV 16 was detected in all cases that were positive by in situ hybridization except for one, which showed HPV 6/11 DNA. In one case there was a mixed triple infection for HPV 6/11, 16, and 18. The patient who was positive for HPV 6/11 had a giant condyloma associated with an inguinal lymph node containing a metastatic well-differentiated squamous cell carcinoma. Three cases were positive for papillomavirus internal capsid species-nonspecific antigen (PV-Ag) (with ultrastructural evidence of virions in one of them) and were negative for HPV-DNA hybridization. They appeared to be infected with a type of HPV not identified by the available probes. Three cases, and two sites of two other cases with double infection, were HPV-DNA-positive and PV-Ag-negative. They illustrate the limitation of immunohistochemical investigation in cases with high-grade intraepithelial neoplasia. Six cases of verrucous carcinoma of the vulva were negative for HPV DNA by in situ hybridization.     

Human papillomavirus DNA sequences in cell lines derived from head and neck squamous cell carcinomas.

There is increasing evidence that human papillomaviruses (HPV) have a casual role in some neoplasms in human beings. As examples, DNA of HPV types 16, 18, and 31 are frequently present in genital cancers in humans. Recently, oncogenic HPV types have also been identified in neoplasms of the head and neck, including verrucous carcinoma of the larynx, squamous cell carcinomas of the oral cavity and larynx, and inverted papillomas of the nose. These findings and our resource of an extensive panel of head and neck squamous cell carcinoma (HNSCC) cell lines led us to begin to investigate how frequently HPV DNA was present in these tumor cell lines. For initial analysis, twenty-two HNSCC cell lines derived from 20 patients' tumors were selected as representative of our tumor cell line panel with respect to diversity of primary site, tumor stage, patient age, sex, and clinical course. For Southern analysis, cell line DNA was tested for hybridization with DNA probes for HPV types 6, 11, 16, 18, and 31. Polymerase chain reaction (PCR) analysis was also performed on five tumor cell lines using types 6, 11, 16, 18, and 52 as probes. Southern blot analysis revealed HPV-specific signals in two of the 22 HNSCC cell lines tested. One of these, UM-SCC-23, was HPV 31 positive, which to our knowledge is the first identification of HPV 31 in HNSCC. UM-SCC-63, the other HPV-positive tumor identified by Southern analysis, hybridized with both type 18 and 31. Of the five tumor cell lines tested with PCR, two were HPV positive.(ABSTRACT TRUNCATED AT 250 WORDS)     

Carcinosarcoma of the uterine cervix: a report of eight cases with immunohistochemical analysis and evaluation of human papillomavirus status

Carcinosarcomas (malignant Müllerian mixed tumors [MMMTs]) of the uterine cervix are rare neoplasms. This report describes the morphology, immunohistochemical profiles, and human papillomavirus (HPV) status of eight cervical MMMTs. Patients' ages ranged from 32 to 93 years (mean, 61 years). Seven cases showed in situ squamous cell carcinoma (SCC). The invasive epithelial component (EC) was composed of combined adenoid basal carcinoma, basaloid SCC, and adenoid cystic carcinoma (ACC) in two cases. Keratinizing SCC, large cell nonkeratinizing SCC, undifferentiated carcinoma, and basaloid SCC predominated in the remaining tumors, one of which had admixed ACC. The sarcomatous component (SC) was homologous and spindled with admixed myxoid areas in three lesions. The ECs and SCs in six MMMTs showed dual immunostaining with epithelial membrane antigen and the pan-keratin marker, MNF116. The SC was vimentin-positive in seven cases. Five tumors had a vimentin-positive EC. The SC was positive for muscle specific actin and/or smooth muscle actin in seven lesions, of which four were desmin-positive. Polymerase chain reaction (PCR) using GP5+/GP6+ L1 consensus primers detected HPV DNA in all eight cases. Nonisotopic in situ hybridization with digoxigenin-labeled probes to HPV types 6, 11, 16, 18, 31 and 33 demonstrated integrated HPV 16 in three cases, not only in the EC, but also in nuclei of the SC. This is the first study to implicate HPV in the evolution of cervical MMMTs. The above observations lend support to a metaplastic theory of histogenesis.     

No evidence of human papillomavirus DNA in breast carcinoma in Tunisian patients

The aim of this study was to evaluate the prevalence of broad range of anogenital HPVs in a series of 123 Tunisian breast carcinoma cases. PCR assays were performed to amplify regions within the L1, E1, E6 and E7 open reading frames of a broad range of anogenital HPVs and specific types HPV16, 18, 31 and 33. In addition, we performed an in situ hybridization analysis using HPV biotinylated DNA probes for the detection of broad spectrum of anogenital HPV types, high-risk HPV types (16 and 18), intermediate-risk HPV types (31 and 33) and low-risk HPV types (6 and 11). None of the 123 breast carcinoma samples showed PCR amplification of HPV DNA using the broad spectrum consensus primer-pairs E1-350L/E1-547R and GP5+/GP6+ primers. Furthermore, neither high risk nor low-risk HPV types were detected in any of these cases. Moreover, using in situ hybridization for the detection of HPVs, we failed to detect a positive signal in neoplastic cells in any case. Our results suggest that anogenital papillomaviruses are unlikely to play a role in the development of breast carcinomas in Tunisian patients.     

Sexually transmitted papilloma viral infection in the male. VII. Is cancer of penis sexually transmitted?

Four cases are reported of squamous cell carcinoma of the genital tract in males. The close association of HPV 16/18 with in situ squamous carcinoma affecting the penis is demonstrated. The first documented case of a primary penile squamous cell carcinoma with metastatic deposits yielding positive HPV 16/18 isolates as shown by in situ DNA hybridization again demonstrates a close association of this virus to malignancy in humans. The clinical implications of these findings strongly suggest that squamous cell carcinoma of the penis is a sexually transmitted disease. A thorough examination of both sexual partners is necessary when Bowenoid papulosis or penile carcinoma is present, since the female partner is at high risk for cervical neoplasia.     

阴茎癌组织中人乳头瘤病毒DNA的原位杂交研究

为了研究人乳头瘤病毒与阴茎癌的关系，应用地高辛标记的ＨＰＶ６、ＨＰＶ１１、ＨＰＶ１６和ＨＰＶ１８ＤＮＡ探针分别对４６例阴茎癌组织进行原位杂交检测ＨＰＶＤＮＡ。结果显示：ＨＰＶＤＮＡ阳性２２例，其中ＨＰＶ１６ＤＮＡ阳性２０例，ＨＰＶ１８ＤＮＡ阳性４例，在２例转移癌和癌旁不典型增生组织中检测到了ＨＰＶ１６ＮＤＡ，未见ＨＰＶ６ＤＮＡ和ＨＰＶ１１ＤＮＡ阳性，ＨＰＶＤＮＡ位于癌细胞核中。结果表明原位杂交可以用来研究阴茎癌组织中ＨＰＶ的存在及分型，同时也证实了阴茎癌的发生和ＨＰＶ１６及ＨＰＶ１８感染有密切关系     

Histological Features of Human Papilloma Virus 16 and Its Association with the Development and Progression of Anal Squamous Cell Carcinoma

Purpose To investigate the development of anal squamous cell carcinoma (SCC) and the expression patterns of human papillomavirus (HPV). Methods We performed typing of HPV-DNA using a polymerase chain reaction (PCR), and amplified the Duchenne muscular dystrophy (DMD) genes simultaneously, to investigate the tumor DNA state. The expression patterns of HPV in the cancer cell nuclei was investigated by in situ hybridization (ISH) using HPV probes. Results Amplification of DMD genes was confirmed in 8 of 20 patients with anal SCC, suggesting that tumor DNA was preserved in these patients. In seven of these eight patients, only HPV16 was detected by both PCR and ISH, suggesting HPV16-induced carcinogenesis. In two patients with carcinoma in situ (CIS), the cancer cells showed only a diffuse pattern (DP), and in two patients with invasive cancer, the cancer cell showed only an oligo-dot pattern (OP). In one patient with lesions ranging from CIS to invasive cancer, the histologic features varied in each area, from DP to OP. This change originated in the deep part of the microinvasive area. Conclusions These findings show that HPV16 infection is closely involved in the development of anal SCC and suggest that the change in the genome occurs at the stage of microinvasive cancer.     

The role of cytology in the diagnosis of benign and malignant anal lesions

Squamous cell carcinoma is a rather infrequent neoplasm of the gastrointestinal tract. Nevertheless its frequency is increasing lately especially in high risk groups of the population infected from HIV or HPV viruses. Squamous cell carcinoma is a slowly and locally growing neoplasm which metastasizes in advanced stages. Its diagnosis must be accomplished by the least traumatic examinations possible. In our study we reviewed our five years experience that included 116 cases. In 89 of them cytological material from ulcerated positions of the anal region was examined. In the rest 27 cytological material was obtained by fine needle aspiration of subcutaneous or submucosal anal lesions. All 116 case reports were retrospectively evaluated. Cytological evaluation revealed 29 cases of normal anal epithelium, 13 granulomas, 12 cases of HPV infection, 28 anal squamous intraepithelial lesions (ASIL), 17 post radiation injuri-es of the anal mucosa and 17 carcinomas. The neoplasms were further subclassified in 12 well differentiated squamous cell carcinomas, 4 cloacogenic carcinomas and 1 leiomyosarcoma. Histological examination followed the initial cytological diagnosis in 75 cases. The correlation between cytological and histological reports did not reveal any false negative or any false positive result. The agreement between histological and cytological evaluation was absolute. Cytological examination is proved to be an easily accessible and totally reliable, low cost diagnostic method, not requiring any kind of anesthesia. It is well accepted by the patients and of paramount clinical utility for the initial diagnostic assessment, the long-term follow up after treatment of anal cancer patients. It is also valuable for the differential diagnosis among benign, premalignant and malignant anal lesions.     

Papillary squamous cell carcinomas of the upper aerodigestive tract: a clinicopathologic and molecular study

BACKGROUND: The limited studies and the small number of published cases of papillary squamous cell carcinoma have precluded accurate assessment of the biologic characteristics of this lesion. METHODS: Thirty-eight of the carcinomas were studied. In-situ hybridization and polymerase chain reaction were performed to detect human papilloma virus (HPV) and p53 expression. RESULTS: HPV was found in 4 of 14 assessable carcinomas by in-situ hybridization and in 5 of 14 by polymerase chain reaction. The most frequently identified HPVs were HPVs in 6/11 and 16/18 patients. In general, a reciprocal relationship was found between p53 and HPV prevalence. The most lethal site for this tumor was the sinonasal tract, whereas patients with papillary squamous cell carcinomas of the larynx had the best outlook. Eleven of 25 (44%) assessable patients died of disease (mean time interval, 2 year). CONCLUSIONS: Papillary squamous cell carcinoma of the upper aerodigestive tract is a distinct variant of squamous cell carcinoma. As such and because of its putative association with HPV, papillary squamous cell carcinoma could be an informative model for defining how viral oncogenes cooperate with other factors in genomic instability, carcinogenesis, and tumor development.     

Papillomavirus found in anorectal squamous carcinoma, not in colon adenocarcinoma.

We used polymerase chain reaction DNA amplification methods for the detection and typing of genital human papillomaviruses in paraffin-embedded tissue sections of five patients with anorectal squamous cell carcinoma and 22 patients with colonic adenocarcinoma. The cases were further tested by in situ hybridization with biotin-labeled probes specific for human papillomavirus types 6/11, 16/18, and 31/33/35. By polymerase chain reaction, human papillomavirus DNA was demonstrated in all of the cases of anorectal squamous cell carcinoma and in none of the cases of colonic adenocarcinoma for which analyzable DNA was available. Tumor cell nuclei stained for human papillomavirus DNA by in situ hybridization in four of the five cases of squamous cell carcinoma and in none of the cases of colonic adenocarcinoma. We conclude that human papillomavirus types usually associated with malignant transformation are uniformly present in anorectal squamous cell carcinoma but are absent from adenocarcinoma of the colon.     

Human papillomaviruses 6/11, 16/18 and 31/33/51 are not associated with squamous cell carcinoma of the urinary bladder

OBJECTIVE: To assess high-risk human papillomavirus (HPV), mainly HPV type 16, 18, 31 and 33 (an important aetiological factor in squamous cell carcinoma, SCC, of the anogenital region) in SCC of the urinary bladder. MATERIAL AND METHODS: Sixteen SCC from the urinary bladder were evaluated using non-isotopic in situ hybridization with a sensitive detection system for the presence of high-risk HPV 16/18, or 31/33/51, and for HPV6/11, a low-risk type commonly found in condylomata. Previously published studies were also reviewed and assessed. RESULTS: No high-risk HPV was found in any of the SCC of the bladder evaluated. Previous reports identified nine HPV-positive SCC of a total of 105, including the present series. In four of these positive cases, HPV types were found that are considered a high risk in anogenital carcinomas. CONCLUSION: From the present and previous results, we conclude that HPV has no major role in the pathogenesis of SCC of the urinary bladder.     

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.     

Association of DNA aneuploidy with human papillomavirus-induced malignant transformation of sinonasal transitional papillomas

The nuclear DNA content of 19 transitional papillomas of the sinonasal region and 9 maxillary squamous cell carcinomas was studied by flow cytometry; the presence of human papillomavirus (HPV) DNA types 11 and 16 was determined by the in situ hybridization technique from paraffin-embedded tissue. Thirteen (68%) of the papillomas and none of the carcinomas contained HPV genome. Six (32%) of the papillomas and 4 (44%) of the carcinomas had an aneuploid DNA content. The relative DNA content (DNA index) of the aneuploid maxillary carcinomas was larger than that of aneuploid papillomas (p = 0.004). Three of the papillomas underwent malignant transformation, all three of which contained HPV type 16 DNA; two were also aneuploid. Data indicate that papillomas containing HPV type 16 DNA have a tendency (p = 0.06) to undergo malignant transformation, and that this tendency is greater if DNA aneuploidy or HPV type 11 DNA is also present (p = 0.02).     

Synchronous and metachronous endocervical and ovarian neoplasms: evidence supporting interpretation of the ovarian neoplasms as metastatic endocervical adenocarcinomas simulating primary ovarian surface epithelial neoplasms

The vast majority of endocervical adenocarcinomas are high-risk human papillomavirus (HPV)-related neoplasms, characterized by p16 expression and frequent loss of hormone receptor expression, which infrequently metastasize to the ovaries. We report 10 cases of endocervical adenocarcinomas with ovarian metastases in which the ovarian tumors simulated primary ovarian surface epithelial neoplasms. The presence of HPV DNA was assessed to determine whether the ovarian neoplasms were metastases or independent neoplasms. Immunohistochemistry for hormone receptors and p16 was also performed. The ovarian metastases presented concurrently with the primary endocervical tumors in 5 cases, subsequent to the endocervical tumors in 3 cases, and prior to diagnosis of the endocervical tumors in 2 cases. The ovarian tumors ranged in size from 2 to 30 cm, with tumors in 7 cases measuring 10 cm or greater. The ovarian tumors were unilateral in 8 cases and bilateral in 2. In all cases, the ovarian tumors were initially diagnosed as or thought to represent independent primary ovarian surface epithelial tumors (atypical proliferative [borderline] tumors or well-differentiated carcinomas of endometrioid or mucinous type). The endocervical tumors ranged in size from microscopic foci to 3 cm, with depth of invasion ranging from 0.2 to 1.5 cm; in 2 cases, the invasive foci qualified as microinvasive according to Federation Internationale de Gynecologie et d'Obstetrique staging criteria for cervical carcinoma. Adenocarcinoma in situ was identified in all tumors. In all cases, the paired endocervical and ovarian tumors contained identical HPV types. All evaluable tumors were diffusely positive for p16; and in 8 cases, there was absent or only limited expression of hormone receptors. Two of the minimally invasive endocervical tumors were initially interpreted as adenocarcinoma in situ and not recognized as unequivocally invasive even when evaluated in conjunction with the histologically identical ovarian tumors. HPV DNA detection in the ovarian tumors of 2 patients without known cervical disease led to discovery of occult cervical adenocarcinomas in those patients. Endocervical adenocarcinomas, including some qualifying as microinvasive, can metastasize to the ovaries and simulate primary ovarian surface epithelial neoplasms. The presence of HPV DNA in these ovarian tumors confirms that they are metastatic endocervical adenocarcinomas.     

Laryngeal Squamous Cell Carcinoma in a 13 Year-Old Child Associated with Human Papillomaviruses 16 and 18: A Case Report and Review of the Literature

Squamous cell carcinoma (SCC) of the larynx is extremely rare in adolescents and typically has an aggressive nature. The mechanism of laryngeal oncogenesis is complex and little is known about the role that human papillomavirus (HPV) plays in SCC in adolescents. We report a case of invasive laryngeal SCC that co-expressed HPV DNA subtypes 16 and 18 in a 13 year-old boy. Detection of HPV DNA types 6, 11, 16, 18, 31, 33, and 51 was performed by in situ hybridization, with confirmation by polymerase chain reaction. Immunohistochemical staining with p16 and HPV 16/18 revealed diffusely positive staining in the tumor cells. Coinfection by HPV DNA types 16 and 18 has not been previously reported, but our case suggests that HPV is a risk factor in developing laryngeal SCC in children and adolescents. Future studies evaluating HPV in the pathogenesis of these lesions is recommended to determine its prognostic significance.