Investigation of human papillomavirus DNA in colorectal carcinomas and adenomas

Human papillomavirus (HPV) has been considered to be an etiological agent for anogenital cancers, such as cervical cancer and possibly a subset of cancers of the aerodigestive tract. The aim of the study was to evaluate the presence of human papillomavirus DNA in colorectal carcinomas and adenomas. Formalin-fixed and paraffin-embedded archival tissue samples were used for DNA extraction. One hundred and six colorectal carcinomas and 62 adenomas were screened by nested polymerase chain reaction (PCR) for HPV DNA with a control group of 49 cervical tissues with invasive cervical carcinoma and cervical intraepithelial neoplasia (CIN). In the study group, we did not find HPV DNA positivity in any of all the colorectal carcinomas and adenomas. In the control group with cervical lesions, 34 out of 49 (69.4%) samples were positive for the HPV DNA. These results indicated that there was no correlation between HPV infection and colorectal carcinomas and adenomas.     

Association of human herpesvirus type 6 DNA with human bladder cancer

We examined the presence of human herpesvirus type 6 (HHV6) DNA in a series of 74 bladder carcinomas from a Mediterranean population to elucidate their possible role as cofactor in the development of bladder cancer with or without associated human papillomavirus (HPV) infection. HHV-6 type B DNA was present in 5 men (6.8%) out of the 74 tumors investigated; two of them had associated HPV-16 DNA in the same specimen. In one case that had associated urothelial carcinoma in situ, both HHV-6B and HPV-16 DNA were present. In conclusion, the low incidence of HHV-6B in bladder cancer and the ubiquitous nature of HHV-6 infection are more consistent with a bystander role rather than cofactor in the oncogenesis of bladder cancer.     

人乳头瘤病毒感染和p53基因突变与宫颈癌的关系

目的：探讨人乳头瘤病毒１６ 和１８ 型及抑癌基因ｐ５３ 突变对宫颈的致癌作用以及 Ｈ Ｐ Ｖ 感染与ｐ５３ 基因突变的相关性。方法：采用聚合酶链反应（ Ｐ Ｃ Ｒ） 技术和限制性酶切片段多态性分析（ Ｒ Ｆ Ｌ Ｐ） 技术对３４ 例原发性宫颈癌组织及３０ 例正常宫颈组织 Ｈ Ｐ Ｖ１６ ,１８ 型 Ｄ Ｎ Ａ 及抑癌基因ｐ５３ 的突变进行了检测。结果： Ｈ Ｐ Ｖ１６ ,１８ Ｄ Ｎ Ａ 在宫颈癌的总阳性率为６４７ ％ （２２／３４） ,正常宫颈组织只有６７ ％ 阳性,８例宫颈癌组织出现ｐ５３ 基因第６ 外显子突变,其中２ 例为 Ｈ Ｐ Ｖ１６ Ｄ Ｎ Ａ 阳性、１ 例 Ｈ Ｐ Ｖ１８ Ｄ Ｎ Ａ 阳性。结论：宫颈癌的发病与 Ｈ Ｐ Ｖ 感染及ｐ５３ 基因突变有关,宫颈癌组织中ｐ５３ 基因突变与 Ｈ Ｐ Ｖ 感染无关。     

Etiologic role of human papillomavirus infection in bladder carcinoma

BACKGROUND:

The authors elucidated an etiologic role of human papillomavirus (HPV) infection in carcinoma of the bladder.

METHODS:

One hundred seventeen of 224 patients with bladder carcinoma who were treated between 1997 and 2009 were enrolled in this study. The presence of HPV DNA was tested on frozen carcinoma tissues that were obtained by transurethral resection using a polymerases chain reaction‐based method. Localization of HPV was observed on archival tissue specimens by in situ hybridization (ISH) for high‐risk HPV DNA. Cyclin‐dependent kinase (CDK) inhibitor 2A (inhibits CDK4) (p16‐INK4a) and minichromosome maintenance protein‐7 (mcm‐7)—surrogate markers for high‐risk HPV‐E7 oncoprotein—and HPV‐L1 (capsid) protein expression were evaluated by immunohistochemistry.

RESULTS:

HPV types 16, 18, 31, 33, 52, and 58, and an unknown HPV type were detected in 18 of 117 samples (15%) from patients with bladder carcinoma. HPV16 was identified in 6 samples, HPV18 was identified in 4 samples, and HPV33 was identified in 3 samples. All were single HPV type infections. HPV was detected in 38% (12 of 28) of histologic grade 1 bladder carcinomas, 8.5% (6 of 71) of grade 2 bladder carcinomas, and in 0% (0 of 18) of grade 3 bladder carcinomas. Multivariate analysis indicated that younger age (<60 years; odds ratio [OR], 10.9; 95% confidence interval [CI], 2.6‐45.3) and grade 1 tumors (OR, 4.5; 95% CI, 1.2‐17.0) were associated with HPV infection. ISH analysis indicated that high‐risk HPV DNA was localized in the nuclei of tumor cells of all HPV‐positive samples. p16‐INK4a and mcm‐7 were expressed in 94% and 89% of HPV‐positive carcinoma cells, respectively. HPV‐L1 protein expression, which suggested reproductive HPV infection, was not observed in any carcinoma.

CONCLUSIONS:

The current results indicated that high‐risk HPV is likely to be a causative agent of some low‐grade bladder carcinomas that develop in younger patients. Cancer 2011. © 2010 American Cancer Society.     

New human papillomavirus sequences in female genital tumors from Japanese patients

Tissues from 52 cervical carcinomas, 15 cervical intraepithelial neoplasias III (CINs III), and 3 vulvar carcinomas were examined for the presence of human papillomavirus (HPV) sequences by Southern blot hybridization with HPV 16 or 18 DNA as the probe. HPV 16 or 18 DNA was detected in only 17 cervical carcinomas (33%), 5 CINs III (33%), and 1 vulvar carcinoma (33%). These frequencies are lower than those reported by others. However, new, as yet unidentified, HPVs were also detected at rather high frequency under less stringent conditions of hybridization using HPV 16 and 18 DNAs as probes. These HPVs did not hybridize with HPV 6, 11, 16, or 18 under stringent conditions, and were different from two other known types of genital tumor-associated HPVs, HPV 31 and HPV 33, judging from the patterns of their restriction enzyme digests. The total frequencies of HPV sequences were 48% (25/52) for cervical carcinomas, 53% (8/15) for CINs III, and 67% (2/3) for vulvar carcinomas.     

Absence of human papillomavirus sequences in epithelial breast cancer in a Mexican female population

The role of human papillomavirus (HPV) in breast cancer is controversial. We evaluated 118 breast carcinomas and two paraffin-embedded tissues of lesions of the nipple of Mexican patients for HPV sequences. No carcinoma sample exhibited koilocytosis, in contrast to lesions of the nipple. We subjected purified DNAs to PCR employing two HPV16/E6 or GP5/6 primer set oligonucleotides. Results showed that HPV DNA sequences were absent in the breast tissues. Absence of HPV in breast carcinoma failed to support an association between HPV infection and this carcinoma.     

The physical state of human papillomavirus type 16 DNA in cervical carcinomas of Hong Kong Chinese.

The presence of human papillomavirus (HPV) in 15 cervical carcinoma specimens obtained from Hong Kong Chinese patients was analyzed by Southern blot hybridization studies. In nine (60%) of them, HPV 16 genomes were detected, while two others (13.3%) were found to harbor HPV DNA of unknown type closely related to HPV 16. All of them were classified as squamous cell carcinomas according to WHO guidelines. In addition, the presence of HPV 18 was shown in another two (13.3%) squamous cell carcinoma samples. Among the nine tumors harboring HPV 16, four specimens (44.4%) have HPV in integrated forms, while four others (44.4%) have HPV in episomal forms. The simultaneous presence of both episomal and integrated forms was demonstrated in the remaining tissue sample (11.2%). The result obtained here indicates a strong association between HPV infection and cervical carcinogenesis in Hong Kong Chinese, with HPV 16 prevalent in squamous cell carcinoma. Moreover, the persistence of HPV 16 episomes in some of the tumor specimens suggests that extrachromosomal HPV DNA, possibly acting synergistically with other oncogenic factors, is also capable of inducing cervical cancer.     

喉癌及其转移淋巴结中检出HPV DNA

应用ＰＣＲ技术对喉原发癌、转移癌、癌旁正常粘膜和声带息肉石蜡包埋组织中的ＨＰＶＤＮＡ进了检测，结果：４０例喉癌标本中２４例检出ＨＰＶＤＮＡ，阳性率６０％（２４／４０），其中ＨＰＶ６／１１、１６／１８ＤＮＡ阳性者分别为６例（５％）和１８例（４５％）；１２例喉癌颈转移淋巴结６例检出ＨＰＶＤＮＡ，这６例均为ＨＰＶ１６／１８；１０例癌旁正常粘膜和１０例声带息肉均阴性。说明喉癌的发生及其淋巴结转移可能与ＨＰＶ１６／１８感染有关。     

Human papillomavirus type 16 and 18 infection is associated with lung cancer patients from the central part of China

Infection with specific high-risk human papillomavirus (HPV) types 16 and 18 have been strongly associated with the genesis of various neoplasms in humans, though such study in lung cancer is limited and the results are controversial. In the present study, we collected and explored 313 fresh lung tumor specimens for the presence of HPV with polymerase chain reaction and non-isotopic in situ hybridization. We found that 44.1% of (138/313) non-small cell lung carcinoma (NSCLC) samples were positive for HPV detection, while 4.2% (4/96) of lung benign controls were positive for HPV 16 and 18 DNA. HPV infection was significant between lung squamous cell carcinoma and adenocarcinoma as well as smoking and non-smoking patients. In HPV-positive lung cancer tissues, abnormal p53 protein accumulation was seen in 97 of the 138 carcinomas (70.3%) and expression of pRb in 54 of the 138 carcinomas (39.1%). There was an obvious relationship between the presence of papilloma viral DNA and abnormal p53 protein accumulation and pRb depletion. Cell proliferation and apoptosis were correlated with HPV infection in NSCLC samples. Our data confirm the high prevalence of HPV in lung carcinomas in the central part of China and suggest the possible mechanism of the carcinogenic role of HPV in these carcinomas.     

HPV infection in cervical-cancer cases in Russia

The presence of human papillomavirus (HPV) sequences in 21 biopsies from cervical carcinomas, II specimens of tissues adjacent to tumours, 2 specimens of cervical tissues with radiation fibrosis from patients after radiation therapy of cervical cancer and 7 normal epithelial tissues from the patients with other genital tumours were examined by polymerase chain reaction (PCR) and Southern-blot analysis. All tumours were HPV-positive by type-specific PCR and 86% by Southern-blot analysis. In normal epithelial and adjacent tissues, HPV sequences were detected in 20% of samples by Southern-blot analysis and in 70% of samples by PCR, including 2 cases of tissues after radiation therapy. HPV 16 was the most prevalent type in tumours (18/21) as well as in normal epithelial tissues (5/7). One HPV-positive tumour contained HPV18 DNA and 2 were doubly infected with HPVs 16 and 18 (2/21). The persistence of exclusively episomal HPV16 DNA was observed in 5 out of 11 tumours examined: 3 cases of squamous-cell carcinomas on the early stage of tumour progression and 2 advanced tumours (squamous-cell carcinoma and adenocarcinoma). The integration of HPV16 genome was detected in 6 out of 11 tumours, but most of them contained episomal forms of viral DNA simultaneously (5 out of 6). The integrative HPV18 genome was found in 2 tumours examined, and the persistence of episomal forms was also observed in one of them. Our data demonstrate that cervical tumours are associated invariably with high-risk types of HPV in Russia. © 1995 Wiley-Liss, Inc.     

Prevalence of human papillomavirus type 16 DNA in cervical carcinoma samples in East Anglia

A substantial increase in the incidence of severely dysplastic cervical lesions (CIN 3) has been observed during the period 1975-1982 in the East Anglian region of England. Since patients with severe dysplasia have an enhanced risk of developing cervical carcinoma, it seems possible that a substantial increase in the rate of cervical carcinoma is likely to occur in the near future. Evidence of a relationship between human papillomavirus (HPV) infection and cervical carcinoma has accumulated recently. We have studied the incidence of HPV16 DNA in cervical tissue samples from patients with cervical carcinoma, severe dysplasias and normal controls. Five out of 11 invasive squamous carcinomas of the cervix, 3/4 dysplasias and 0/12 normal samples were positive in Southern blot assays for HPV16 DNA. Some of the tissue samples had as many as 500 copies of HPV16 DNA per cell. The amount of HPV16 DNA present correlated with the aggressiveness of tumour growth.     

High frequency of human papillomavirus infection in carcinoma of the urinary bladder.

BACKGROUND AND METHODS. The prevalence of type 6, 11, 16, 18, and 33 human papillomavirus (HPV) was investigated with the polymerase chain reaction (PCR) on formalin-fixed, paraffin wax-embedded material, including 48 neoplastic and 21 normal urinary bladder specimens. The PCR-amplified DNA were analyzed by gel electrophoresis and dot blot and Southern blot hybridization. Some tissues were tested further by nonisotopic in situ hybridization. RESULTS. HPV DNA was detected in 39 (81%) of 48 carcinomas and 7 (33%) of 21 normal urinary bladder specimens. The presence of high-risk HPV (types 16, 18, and 33) was increased significantly in carcinoma cases (62%) as compared with normal specimens (14%) (P less than 0.01). Similarly, multiple HPV infections were significantly higher in carcinoma (60%) than in the normal tissues (5%) (P less than 0.01). The overall and high-risk HPV infections in both neoplastic and normal specimens were distributed almost equally in male and female patients. There was no significant correlation between positive results for HPV and histologic grades of the carcinoma. CONCLUSIONS. These results demonstrate that the urinary bladder in both sexes is another site where infection with the common genital tract HPV may carry a risk of malignant transformation.     

Detection and typing of human papillomavirus in archival cervical cancer specimens by DNA amplification with consensus primers

We developed a polymerase chain reaction DNA amplification system using two distinct consensus oligonucleotide primer sets for the improved detection and typing of a broad spectrum of human genital papillomavirus (HPV) sequences, including those of novel viruses. The system incorporates one primer set designed to amplify a highly conserved L1 domain and a second primer set designed to amplify a domain within the E6 gene. We used this system to analyze 48 fixed, paraffin-embedded tissue sections (41 specimens from 33 cervical carcinomas, four normal cervical tissues, and several control tissues) for the presence of HPV DNA. HPV sequences were detected in all carcinoma samples and none of the control samples. Hybridization analyses showed that the results obtained with the two amplification schemes concurred completely. This approach allowed rapid confirmation of typing results and may improve the likelihood of detecting a wide variety of HPV sequences, including those of novel HPVs.     

Histological types of carcinoma of the uterine cervix and the detectability of human papillomavirus DNA

Using the Southern DNA hybridization technique, tissues from 17 cases of invasive carcinoma of the uterine cervix, including nine cases of squamous cell carcinoma, four cases of adenocarcinoma, one case of adenosquamous carcinoma, and three cases of undifferentiated carcinoma, were examined for the presence of human papillomavirus (HPV) DNA. None of the studied cases had histologically confirmed association of condyloma acuminatum or cervical intraepithelial neoplasia in the vicinity. HPV DNA was detected in two of 17 cases under low stringency conditions. One lesion was undifferentiated carcinoma, and another was squamous cell carcinoma. Hybridization under high stringency conditions with a variety of HPV DNA probes indicated the presence of HPV-16 in these two lesions. The other HPV-positive lesion was adenocarcinoma, demonstrating weak hybridizations with HPV-2 and HPV-16 DNA probes only under high stringency conditions. Altogether, three of 17 cases (17.6%) contained HPV DNA. This observation contrasts to the rate of HPV DNA present in 15 of 18 cases (83.3%) of the tissues of cervical intraepithelial neoplasia. Our data suggest that HPV was not consistently detected in invasive squamous cell carcinoma, despite the frequent association of HPV with its supposed precursor lesions of cervical intraepithelial neoplasia.     

Genital papillomavirus infection after treatment for cervical intraepithelial neoplasia (CIN) III

Genital human papillomavirus (HPV) infection was studied in 150 women after conization for cervical intraepithelial neoplasia grade three (CIN III). Colposcopically directed biopsies were taken from the cervix and vulva for histopathological diagnosis. 77 specimens were further analyzed immunohistochemically for the presence of HPV capsid antigen. In ten randomly selected cervical biopsies cellular DNA was dot blot hybridized with HPV 6/11 and 16/18 DNA probes. Genital warts were seen in 10 (7%) of the patients. Among the routine cytological smears, HPV infection was only reported in 3 (2%). In 87/142 (61%) of the cervical tissues koilocytes were found. A further 9/142 (6%) associated with CIN. Of the vulvar biopsies 91/145 (63%) contained koilocytes. A further 12/145 (8%) were associated with vulvar intraepithelial neoplasia. HPV capsid proteins were detected in 35/77 (45%) of the cervical and in 20/41 (49%) of the vulva biopsies. All cervical DNA samples hybridized with probes for HPV 6/11 and two also reacted with the HPV 16/18 probes. Conclusion: A latent HPV infection of the cervix or vulva, can be detected in 85% of the women previously treated for CIN III by conization.     

Telomerase activity and human papillomavirus (HPV) infection in human uterine cervical cancers and cervical smears

Telomerase activity and human papillomavirus (HPV) infection were investigated in uterine cervical samples using molecular biology techniques. Thirteen cervical carcinomas and corresponding normal tissue from the same patient, and 102 cervical swabs were examined. Telomerase activity was detected in 12 of 13 cervical cancer tissues (92%). Of the 12 cases that showed telomerase activity, all were HPV positive, and the one case that did not show telomerase activity was HPV negative. A telomeric repeat amplification protocol assay detected telomerase activity in one out of seven normal cervical tissues (14%), and this one case was HPV positive. In cervical smear samples, telomerase activity was detected in two out of 36 normal smears (6%; both HPV positive), in 10 of 32 (31%) CIN1 (cervical intra-epithelial neoplasia) cases (three HPV positive), in four of five (80%) CIN2 cases (two HPV positive), in 15 of 21 (71%) CIN3 cases, (seven HPV positive) and in seven of eight (88%) squamous cell carcinoma cases (six HPV positive). These results suggest that telomerase activity may play some role in cervical carcinogenesis, and telomerase activity is associated with HPV infection in uterine cervical lesions.     

Oncogenic association of specific human papillomavirus types with cervical neoplasia

Molecular hybridization analysis of human papillomavirus (HPV) DNA from 190 cervical biopsy specimens from women in the United States, Brazil, and Peru revealed viral sequences in 2 (9%) of 23 biopsy specimens of normal mature squamous epithelium, 7 (44%) of 16 biopsy specimens of metaplastic squamous epithelia, 60 (77%) of 78 cervical intraepithelial neoplasia (CIN), 57 (89%) of 64 invasive squamous carcinomas, and 8 (89%) of 9 endocervical adenocarcinomas. HPV typing by DNA hybridization revealed HPV 6 and HPV 11 sequences in metaplastic squamous epithelia, CIN I, and CIN II, but not in CIN III lesions or invasive carcinomas. HPV 16 was detected in metaplastic epithelium and in nearly half of the invasive squamous carcinomas and adenocarcinomas. It was present in 31% of CIN lesions, increasing in frequency with the severity of CIN from 20% of CIN I to 50% of CIN III. HPV 16 showed a striking difference in geographic distribution, being detected in 36% of the carcinomas from the United States compared to 64% of the carcinomas from Brazil and Peru. HPV 18 was found in metaplastic epithelia and in 17% of carcinomas but in only 1% of CIN lesions. HPV 31 was not found in metaplastic epithelium but was present in 6% of carcinomas and in 18% of CIN lesions. In addition, a group of uncharacterized HPVs, not corresponding to any of the probes used, was found in 5% of normal and metaplastic epithelia and in 18% of CIN and 19% of invasive cancers. These results suggest that individual HPV types that infect the cervix have varying degrees of oncogenic association. HPV 6 and HPV 11 appear to have very little oncogenic association, HPV 31 has low oncogenic association, and HPV 16 and HPV 18 have high oncogenic association.     

Occurrence of human papillomavirus types 16 and 18 DNA in cervical carcinomas from Japan: age of patients and histological type of carcinomas

We have detected by DNA hybridization human papillomavirus (HPV) types 16 and 18 DNA sequences in 19 and 3, respectively, out of 56 cervical carcinomas from Japan. Eighteen out of 19 HPV 16-positive specimens were from squamous cell carcinomas, whereas the three HPV 18-positive specimens were from a squamous cell carcinoma (1/50), an adenosquamous carcinoma (1/3), and an adenocarcinoma (1/3). The occurrence of HPV 16 DNA decreased in patients over 60 years old (less than or equal to 60 years, 15/34 (44%); 60 years less than, 4/22 (18%)) (P less than 0.05).