E6/E7 expression of human papillomavirus types in cutaneous squamous cell dysplasia and carcinoma in immunosuppressed organ transplant recipients (original) (raw)
Related papers
Journal of Investigative Dermatology, 2005
Epidermodysplasia verruciformis (EV) type human papillomavirus (HPV) DNAs have been detected by PCR in squamous cell carcinomas (SCCs) from both organ transplant recipients (OTR) and immunocompetent (IC) individuals. Their role in the development of skin cancer remains unclear, and previous studies have not addressed whether the viruses are transcriptionally active. Here we have used in situ hybridisation (ISH) to investigate the transcriptional activity and DNA localisation of these viruses. EV-HPV gene transcripts were demonstrated in 4 of 11 (36%) OTR SCC, 1 of 2 (50%) IC SCC and 1 of 5 (20%) OTR warts positive by PCR. Where detected, viral DNAs co-localised with E2/E4 early region gene transcripts in the middle or upper epidermal layers. Non-EV cutaneous HPV gene transcripts were demonstrated in 1 of 5 (20%) OTR SCC and 4 of 10 (40%) OTR warts. Where mixed infection was present by PCR, transcripts for both types were detected in 2 of 6 (33%) cases. Our results provide evidence of EV-HPV gene expression in SCCs. Although only a proportion of tumours were positive, the similarly low transcriptional activity in viral warts suggests that this may be an underestimate. These observations, together with emerging epidemiological and functional data, provide further reason to focus on the contribution of EV-HPV types to the pathogenesis of cutaneous SCC.
Volume 4(5): 1-5 types that taxonomically correspond to A7 (HPV 18, 39, 45, 59 and 68) and A9 species (HPV 16, 31, 33, 35, 52 and 58) include most of the so called high risk types, being types 16 and 18 responsible for about 60-75% of all precursor lesions and squamous invasive cancers Abstract Persistent high risk Human Papillomavirus (HPV) infection is necessary for the development of cervical cancer (CC). HPV carcinogenesis is based on viral E6 and E7 proteins' capacity to interfere in cell proliferation control. The metastasis status of pelvic lymph nodes (PLN) is a critical parameter in post-operative decisions on adjuvant therapy, given its strong correlation with recurrence in CC. In order to complement the histopathological evaluation of subclinical node metastases, we evaluated the application of a commercial HPV E6 / E7 mRNA kit, to detect viral messenger RNAs in lymph nodes and tumors of patients with CC. Forty five cervical primary tumors and 152 PLN (3-4 from each patient) were included. HPVs were typed in tumors by PCR (polymerase chain reaction) using generic primers PGMY and reverse line blot hybridization (RLB) with type-specific oligo probes corresponding to 37 HPV types that infect the anogenital tract. PNL were collected in RNAlater (Invitrogen) and mRNA was extracted using the MiniMag (Biomerieux) system. Detection of E6 and E7 mRNAs corresponding to HPVs types 16, 18, 31, 33 and 45 was carried out using isothermal real time PCR (NucliSENS EasyQ HPV, Biomerieux). HPV was detected in the tumors of 42 patients; the viral types identified were HPV16 (n = 32), HPV18 (n = 5), HPV31 (n = 3), HPV45 (n = 2), HPV59 (n = 1) and HPV73 (n = 1). These last 2 cases were not considered in the study because the mRNA detection system does not include these viral types. Also excluded were 2 cases whose tumors were HPV negative. There was a high correlation between the histological and virological results. Seventy-two percent of the histologically positive PLN were positive for E6-E7 mRNA; while 93% (125/134) of the negative PLN were also negative for E6-E7 mRNA assay. There was, however 8% (9/134) of negative PLN in which viral messengers were detected. Patients' follow up was limited (4 years) and except for two patients who died (both positive for HPV 45) in the period under review, no recurrences were recorded in any of the patients included in the study. The presence of HPV in PLN may indicate a metastasis, as since the virus is not able of producing viremia or invading tissues it can only be "transported" by the cancer cell. The presence of HPV mRNA indicates viral genome transcription, a process which occurs in a lymph node only in CC metastatic cells, as these viruses can only replicate in epithelial-origin cells. Therefore, the findings of these mRNAs in the negative pelvic lymph nodes point out a very early metastasis, detectable at molecular level, but unobservable on histological diagnosis. The commercial kits for HPV E6/E7 mRNA detection, usually applied for the cervical disease management, may be used also to evaluate PLN biopsies, in hospitals without high complexity laboratories. The test might be a tool to complement morphological observation, particularly when PLN do not exhibit characteristics compatible with invasion, thus optimizing monitoring and decision making.
Experimental and Molecular Pathology, 2017
Development of cutaneous carcinomas has been associated with HPV infection. There have been various reports on p16, p53 and pRb expression in cutaneous carcinomas and on its linkage to HPV status. Association of protein expression and HPV infection with DNA content is not clear. The aim of this study was to determine a possible correlation between HPV type, protein expression and DNA content in both pre-invasive and invasive squamous cell carcinoma, as well as differences between studied groups in these parameters. Sections of formalin fixed paraffin-embedded tumor tissue from 54 cases of Morbus Bowen (preinvasive cutaneous carcinoma) and 41 cases of invasive squamous cell carcinoma of the skin were subjected to HPV genotyping using Lipa (Line imuno probe assay), immunohistochemical staining for p16(INK4A), p53, pRb and prepared for Flow cytometry DNA content analysis. Obtained data were analyzed in SPSS using Chi square test. Only p16 expression showed statistically significant differences in studied groups. Statistically significant correlations were found only in MB between parameters HPV-p53, p53-pRb and p53-p16. Our results suggest different virus-induced pathobiology pathways for different cutaneous carcinoma groups.
Journal of Clinical Microbiology, 2006
The oncogenic potential of the human papillomavirus (HPV) early genes E6 and E7 is well established and a source of interest with regard to HPV testing for cervical carcinoma. Here we present a study performed with 204 histologically confirmed invasive cervical squamous cell carcinomas (SCCs) in which we evaluated the HPV E6 and E7 mRNA detection assay PreTect HPV-Proofer for detection of high-risk HPV types 16, 18, 31, 33, and 45. For further evaluation, detection of E6 and E7 mRNA from HPV types 35, 52, and 58 by real-time multiplex nucleic acid sequence-based amplification was also included. For comparison and to assess the overall prevalence of various HPV types, samples were also tested for HPV DNA by both consensus and type-specific PCR, reverse line blotting, sequencing, and in situ hybridization. The overall prevalence of HPV was 97%. HPV E6 and E7 transcripts were detected in 188 of 204 (92%) biopsy specimens, of which 181 contained one of the following HPV types: 16, 18, 31, 33, or 45. Consensus PCR and type-specific PCR detected HPV in 187 of 204 and 188 of 204 (92%) specimens, respectively. In conclusion, this study verifies the presence of HPV E6 and E7 mRNA in SCCs and demonstrates that HPV infections among Norwegian women with SCCs are limited mainly to the five high-risk types, 16, 18, 31, 33, and 45. This, together with the fact that PreTect HPV-Proofer detects the HPV oncogenic transcripts, suggests that the assay is a valuable approach in the field of HPV detection in cervical carcinoma.
Journal of Investigative Dermatology, 2008
The presence of certain types of human papillomavirus (HPV) is a known risk factor for the development of anogenital squamous cell carcinomas (SCCs). A similar association has been hypothesized for cutaneous SCCs, although, to our knowledge, no studies to date have combined sensitive HPV DNA detection techniques with epidemiologic data controlling for known risk factors to explore the association. We designed a case-control study examining HPV prevalence using highly sensitive PCR-detection assays in tissue samples from 85 immunocompetent patients with histologically confirmed SCCs and 95 age-matched individuals without a prior history of skin cancer. A standardized interview was administered to all study subjects to collect information pertaining to potential confounding variables. The overall detection rate of HPV DNA was high in case lesions (54%) and perilesions (50%) and in both sun-exposed normal tissue (59%) and nonsun-exposed normal tissue (49%) from controls. In comparing case tissue to control tissue, there was no differential detection of HPV DNA across various HPV species. However, HPV DNA from β-papillomavirus species 2 was more likely to be identified in tumors than in adjacent healthy tissue among cases (paired analysis, odds ratio = 4.0, confidence interval = 1.3-12.0). The high prevalence of HPV DNA detected among controls suggests that HPV DNA is widely distributed among the general population. However, the differential detection of HPV βpapillomavirus species in tumors among cases suggests that certain HPV types may be involved in the progression of cutaneous SCCs.
Cancer research, 1994
A total of 118 biopsies from skin lesions of 46 renal allograft patients was analyzed for human papillomavirus (HPV) DNA by polymerase chain reaction with degenerate primers and also partially by subsequent se quencing of the amplified fragment. Sixty-two % of the benign prolifer ations (31 of 50) contained DNA of known HPV types as well as HPV sequences related to a number of epidermodysplasia verruciformis-associated HPV types. HPV DNA sequences were found in 14 (56%) of 25 biopsies from squamous cell and basal cell carcinomas. One squamous cell carcinoma contained HPV 41 DNA. A novel 640-base pair fragment sharing homology with HPV 29 (82.7%) was found in 15% (3 of 20) of squamous cell carcinomas, in 9.4% (3 of 32) of dysplastic warts and in 8.5% (4 of 47) common warts. The remaining positive carcinoma biopsies contained HPV-related DNA in such a low copy number that additional analysis is required. The identification of new HPV types in skin cancers of immunosuppressed patients (other than epidermodysplasia verruciforniis patients) further expands the spectrum of HPV-linked human malig nancies and permits new approaches to study the pathogenesis of skin cancers.
A total of 118 biopsies from skin lesions of 46 renal allograft patients was analyzed for human papillomavirus (HPV) DNA by polymerase chain reaction with degenerate primers and also partially by subsequent se quencing of the amplified fragment. Sixty-two % of the benign prolifer ations (31 of 50) contained DNA of known HPV types as well as HPV sequences related to a number of epidermodysplasia verruciformis-associated HPV types. HPV DNA sequences were found in 14 (56% ) of 25 biopsies from squamous cell and basal cell carcinomas. One squamous cell carcinoma contained HPV 41 DNA. A novel 640-base pair fragment sharing homology with HPV 29 (82.7%) was found in 15% (3 of 20) of squamous cell carcinomas, in 9.4% (3 of 32) of dysplastic warts and in 8.5% (4 of 47) common warts. The remaining positive carcinoma biopsies contained HPV-related DNA in such a low copy number that additional analysis is required. The identification of new HPV types in skin cancers of immunosuppressed patients (other than epidermodysplasia verruciforniis patients) further expands the spectrum of HPV-linked human malig nancies and permits new approaches to study the pathogenesis of skin cancers.
Journal of The National Cancer Institute, 1996
Background: Nonmelanoma carcinomas of the skin represent the most frequent cancers among the Caucasian population worldwide. They occur with high frequency in renal allograft recipient patients after prolonged immunosuppression. Purpose: We analyzed tumors obtained from both immunosuppressed and nonimmunosuppressed patients for human papillomavirus (HPV) DNA. Methods: Twenty-nine specimens of nonmelanoma carcinomas of the skin were obtained from 19 renal allograft recipient patients; these included 20 specimens of squamous cell carcinoma (SCC) from 11 patients, five specimens of basal cell carcinoma (BCC) from four patients, and four specimens of carcinoma in situ (CIS) from four patients. Forty-one specimens of nonmelanoma carcinomas of the skin were obtained from 32 nonimmunosuppressed patients; these included 26 SCC specimens from 19 patients, 11 BCC specimens from nine patients, and four keratoacanthoma (benign epithelial tumor) specimens from four patients. A polymerase chain reaction method involving use of degenerate oligonucleotide primers, in which the conserved region of the open reading frame of the HPV LI (major capsid protein) gene is amplified, was used to amplify total cellular DNA purified from individual tumors. The DNA of each specimen was subjected to 16 different amplification reactions; different primer combinations were used in order to increase the sensitivity and specificity of HPV detection. Resulting products were probed with a radioactively labeled, degenerate oligonucleotide. HPV-specific DNA was either sequenced directly after elution from the gel or amplified with semi-nested, degenerate primers, after which the products were cloned and sequenced. Sequences were compared with all known papillomavirus sequences. Results: Thirteen (65%) of the 20 SCC specimens and three of the five BCC specimens from immunosuppressed (renal allograft recipient) patients contained identifiable HPV-related sequences, among them 13 putative novel HPV genomes. In addition, all other malignant tumor specimens from this patient group revealed faint signals upon amplification and hybridization; the origin of these signals has not been identified in the present study. In nonimmunosuppressed