宫颈癌细胞中人乳头瘤病毒感染对DNA含量及细胞周期的影响

目的探讨人乳头瘤病毒（ＨＰＶ）感染在宫颈癌变过程中的作用。方法应用聚合酶链反应（ＰＣＲ）和流式细胞计量术（ＦＣＭ）对３９例宫颈癌标本中ＨＰＶ１６和１８型感染、ＤＮＡ含量和细胞周期分布进行检测和分析。结果３１例鳞癌中ＨＰＶ１６阳性１１例（３５．５％），ＨＰＶ１８阳性１例；７例腺癌中ＨＰＶ１６全部阴性，ＨＰＶ１８阳性２例（２８．６％）；１例腺鳞癌ＨＰＶ１６和１８均阴性。总的异倍体率为５９％，二倍体率为４１％。鳞癌异倍体率为７１％（２０／３１），腺癌为４３％（３／７）。鳞癌中ＨＰＶ１６阳性组与阴性组间ＤＮＡ指数和Ｓ期比率有明显差异，而异倍体率无差异。结论ＨＰＶ感染与宫颈癌发生有关。     

Estrogen receptor localization in normal and neoplastic epithelium of the uterine cervix

To investigate the estrogen receptor (ER) status of cells during carcinogenesis of the uterine cervix, the immunohistochemical reactivity for a monoclonal anti-ER antibody (H 222) was studied in 26 normal cervical specimens, 21 cases of cervical intraepithelial neoplasia (CIN), and 21 cases of invasive cervical carcinoma. In addition, the presence of human papillomavirus (HPV) DNA (types 6/11, 16/18, or 31/33/35) was analyzed by in situ hybridization. In the normal cervix, basal cells of the squamous epithelium, metaplastic cells, and endocervical glandular cells were ER positive. In contrast, neoplastic cells of CIN (17 of 21 cases) and invasive carcinoma (19 of 21 cases) were ER negative. The remaining four cases of CIN and two cases of invasive carcinoma were focally ER positive. The HPV DNA analysis revealed that HPV DNA in ER-negative cases was either types 16/18 or undetectable, but all ER-positive neoplasms contained HPV DNA types 31/33/35. These results suggest that most neoplastic cells in CIN and invasive cervical carcinoma lose their ER expression and that this may be related to the HPV DNA types which they possess.     

HPV感染的形态特征及HPV型别与宫颈癌的临床病理联系

目的观察人乳头瘤病毒（HPV）感染的形态学改变,分析挖空细胞形态、HPV型别与宫颈鳞状细胞癌临床病理参数间的联系。方法对594例宫颈鳞状细胞癌进行了形态学观察,按照WHO标准进行组织学分型及分级。运用SPFIO—PCR技术进行HPVDNA扩增,并采用DEIA（DNA enzyme immunoassay）及LiPA（line probe assay）方法进行DNA检测及分型。结果经检测HPVDNA阳性者581例,其中394例具有HPV感染的组织学特点,即出现典型（104例）或不典型（290例）的挖空细胞,检出率为67．8％。36．2％的角化型及72．2％的高分化宫颈鳞状细胞癌中可见典型挖空细胞,而34．0％的非角化型及50．7％的低分化者中无挖空细胞。在HPV型别检测中,以HPV16阳性最多见为453例,其中高、中、低分化者分别为16例、389例及48例;其次为HPV18共43例,其中中分化者32例,低分化者11例;其他高危型HPV按照阳性数多少依次为HPV31、52、59、58、39、45、33、56、66、68、73,分别为17例、12例、12例、11例、6例、5例、5例、3例、2例、1例、1例;低危型HPV包括6及53各1例,且均同时伴有高危型HPV感染。结果显示,HPV16、HPV18阳性者较其他高危型HPV阳性者发病年龄轻;HPV18阳性者较HPV16阳性者分化程度低;HPV18阳性者较HPV16阳性者无挖空细胞病例所占比例较高（分别为48．8％及31．8％）;患者年龄越大FIGO分期越晚。结论挖空细胞形态与宫颈鳞状细胞癌的分化程度密切相关,典型挖空细胞多出现在分化较成熟的癌组织中,而分化较差者常无挖空细胞。HPV16、18是宫颈鳞状细胞癌最重要且最危险的HPV型别。HPV18阳性可能提示宫颈鳞状细胞癌恶性程度高。     

HPV types and cofactors causing cervical cancer in Peru

We conducted a hospital-based case-control study in Peru of 198 women with histologically confirmed cervical cancer (173 squamous cell carcinomas and 25 cases of adenocarcinoma/adenosquamous carcinoma) and 196 control women. Information on risk factors was obtained by personal interview. Using PCR-based assays on exfoliated cervical cells and biopsy specimens, HPV DNA was detected in 95.3% of women with squamous cell carcinoma and in 92.0% of women with adenocarcinoma/adenosquamous carcinoma compared with 17.7% in control women. The age-adjusted odds ratio was 116.0 (95% Cl = 48.6-276.0) for squamous cell carcinoma and 51.4 (95% Cl = 11.4-232.0) for adenocarcinoma/adenosquamous carcinoma. The commonest types in women with cervical cancer were HPV 16, 18, 31, 52 and 35. The association with the various HPV types was equally strong for the two most common types (HPV 16 and 18) as for the other less common types. In addition to HPV, long-term use of oral contraceptives and smoking were associated with an increased risk. HPV is the main cause of both squamous cell carcinoma and adenocarcinoma in Peruvian women.     

Detection of human papillomavirus DNA in esophageal carcinoma in Japan by polymerase chain reaction.

BACKGROUND. Human papillomaviruses (HPV) have been implicated strongly in the pathogenesis of human squamous cell carcinomas, especially of anogenital carcinomas. Some pathologic changes of the esophagus may be one of the candidates for HPV etiology, but the role of HPV infections in the carcinogenesis of the esophagus remains to be clarified. METHODS. To elucidate the association of HPV with carcinogenesis of the esophagus, 45 biopsy samples of esophageal squamous cell carcinomas were examined for the presence of HPV DNA by polymerase chain reaction (PCR). Primers for PCR were (1) consensus primers (CP) for the simultaneous amplification of the E6-E7 regions of cancer-associated HPV types (HPV 16, 18, 31, 33, 52b, and 58), which have been shown to have transforming activities; (2) type-specific primers (SP16, SP18) for the E7 regions of HPV 16 and HPV 18, respectively; and (3) general primers (GP) for the simultaneous amplification of the L1 regions of HPV 6, 11, 16, 18, 31, and 33. RESULTS. PCR using CP first was done for screening and showed that 3 (6.7%) of 45 specimens contained HPV 16 or HPV 18 DNA, the oncogenic high-risk HPV types. This was confirmed by SP16 and SP18 PCR. However, no HPV DNA was detected by PCR using GP. These results suggested that the HPV DNA detected might be integrated into the cell genome with their transforming genes retained and their late regions deleted. CONCLUSIONS. Most oncogenic types of HPV (HPV 16 and HPV 18) were detected by PCR in carcinomas of the esophagus. Thus, HPV might play a role, although at a low frequency, in carcinogenesis of the esophagus.     

Does human papilloma virus have a role in squamous cell carcinoma of the colon and upper rectum?

AIM: To determine if Human Papilloma Virus (HPV) has a role in the aetiology of adenosquamous and squamous cell carcinoma of the colon and upper rectum, and to describe the clinical features seen in our patients with this condition.METHODS: Patients were identified with squamous cell carcinoma (SCC), adenosquamous carcinoma (Ad-SCC), or adenocarcinoma with squamous metaplasia (AA) of the colon and upper rectum over the 10 years from 1/1/1990 to 31/12/1999. Patients were identified from a prospective pathology database. All tumours were at least 5cm above the dentate line. Pathology blocks were stained using the Peroxidase labelled Streptavidin technique using mouse monoclonal antibody NCL-HPV-4C4, which identifies HPV 6, 11, 16 and 18. Age, gender and site matched controls (colorectal adenocarcinomas) were also stained. The clinical presentation and management was reviewed from the case notes.RESULTS: Twenty patients were identified from a pathological database of 2351 colorectal cancers (0.85% of colorectal cancers). 0/20 of the study patients (SCC, Ad-SCC, AA) or adenocarcinoma controls stained positively for HPV 6, 11, 16, 18. The clinical presentation was similar to patients presenting with adenocarcinomas.CONCLUSIONS: The peroxidase labelled streptavidin technique is an immunohistochemical technique with high specificity but lower sensitivity. There was no apparent association between HPV 6, 11, 16, 18 and squamous cell and adenosquamous carcinoma of the colon and rectum using this technique. Clinical features are similar in squamous and adenosquamous colorectal carcinomas to colorectal adenocarcinomas.     

Human papillomavirus type 31 DNA detected in part of the dysplasia but in no part of the squamous metaplasia in a specimen taken from one patient.

Using in situ hybridization, human papillomavirus (HPV 6, 16, 18, 31, 33) DNAs were detected in a cervical severe dysplasia accompanied by squamous metaplasia. It was found that, only HPV 31 DNA was harbored in the cervical severe dysplasia, but HPV DNAs were not identified in a lesion of squamous metaplasia. The in situ hybridization method will be of use, therefore, when dysplasia with squamous metaplasia or other lesions are examined for HPV DNA. In a cervical smear, HPV 31 DNA could be detected on the nuclei of dysplastic cells, so this method is applicable to cervical smears. If squamous metaplasia is to be considered as a precursor lesion to cervical dysplasia, the HPV DNA harbored in the dysplasia must also be detected in the accompanying squamous metaplasia. Our results suggested that not all squamous metaplasias were involved with HPV, as far as we were able to detect using five types of HPV DNA probe.     

Human papillomavirus in lung carcinomas among three Latin American countries

The presence of human papillomavirus (HPV) genome in lung carcinomas has been reported worldwide but its frequency varies from country to country. We examined HPV genome in 36 lung carcinomas, consisting of 14 squamous cell carcinomas, 13 adenocarcinomas, and 9 small cell carcinomas, collected from Colombia, Mexico and Peru. PCR analysis using GP5+/GP6+ primers, combined with Southern blot hybridization, found the presence of HPV genome in 10 (28%) of 36 cases. This percentage is similar to the value of 22% reported by Syrj?nen, who conducted a meta-analysis of nearly 2500 lung carcinomas examined to date. Genotype analysis revealed that the most predominant genotype was HPV-16 (7 cases), followed by HPV-18 (2 cases) and HPV-33 (1 case). HPV-16 was more frequently found among female than male cases (P=0.008) but was not detected in any adenocarcinoma cases. On the other hand, HPV-18 and HPV-33 were detected only among male cases. These HPV genotypes were detected only in adenocarcinomas, and all the HPV genotypes detected in this histological type were HPV-18 or HPV-33. The frequency of HPV-16 positive cases among all the HPV positive cases differed in the sexes (P=0.033) and differed in the three histological types (P=0.017). The presence of HPV tended to be more frequent in well-differentiated tumors when squamous cell carcinomas and adenocarcinomas were combined. However, it was not statistically significant (P=0.093). Neither p16 nor p53 expression in carcinoma cells was related to the proportion of HPV-positive cases. In conclusion, high-risk HPV DNA was detected in 28% of lung carcinomas. The predisposition of HPV-16 to female cases and to non-adenomatous carcinomas warrants further investigation.     

Human papillomavirus DNA in normal, metaplastic, preneoplastic and neoplastic epithelia of the cervix uteri

Colposcopically directed cervical punch biopsies from 362 patients were screened by Southern blot hybridization for the presence of DNA of human papillomavirus (HPV) 6, 10, 11, 16, 18, 31 and 33. The biopsies represented original squamous epithelium, epithelium of metaplastic origin, different stages of cervical intraepithelial neoplasia (CIN) and invasive carcinomas. HPV6/11, 16, 18 and 31 were detected in 2.9% to 13.7% of histologically normal epithelia. HPV6/11 prevailed in CIN I. HPV16 was clearly more abundant than other HPV types in high-grade CIN and invasive cancers (50%-60%), compared with healthy epithelium. Restriction enzyme cleavage analysis of DNA from primary cancers and corresponding metastases proved the stable association of HPV16 DNA with invasive tumor cells. Preliminary follow-up studies of CIN II patients suggested that HPV16-associated lesions are relatively more likely to persist or to progress. Taken together, the data support the notion of a higher oncogenic potential of HPV16.     

采用原位杂交法检测生殖系统尖锐湿疣和鳞状细胞癌内乳头瘤病毒的感染

目的　探讨HPV感染与生殖系统尖锐湿疣和鳞状细胞癌的关系。方法　采用HPV6 /11、16 /18原位杂交试剂盒 ,对 5 5例生殖系统不同病变中乳头瘤病毒感染状况进行分析检测。结果　 2 0例生殖系统尖锐湿疣 19例阳性 ,其中 17例为HPV6 /11型 ,2例为HPV16 /18型。 2 0例生殖系统鳞状细胞癌中 ,15例阳性 ,均为HPV16 /18型。 10例正常生殖系统鳞状上皮组织和 5例外阴白斑组织均呈阴性。结论　HPV6 /11多与生殖系统良性疣状病变有关 ,而HPV16 /18感染多见于生殖系统恶性病变。     

HPV Positive Bronchopulmonary Carcinomas in Women with Previous High-grade Cervical Intraepithelial Neoplasia (CIN III)

A significant higher incidence of some cancers, especially lung cancer, has been found in women with previous HPV-related (human papillomavirus) urogenital and anal neoplasias than in individuals without this particular clinical history. The aim of our study was to investigate whether HPV is present in both CIN III (cervical intraepithelial neoplasia) lesions and bronchopulmonary second primary cancers in women with a clinical history of both diseases. Paraffin-embedded tumour tissue from 75 patients with bronchopulmonary carcinomas was examined using the polymerase chain reaction (PCR) technique and in situ hybridization for the presence of human HPV. In total, 51 primary tumours without metastases, 11 primary tumours with metastases and 13 lymph node metastases without available tissue from primary tumours were analysed. In our study 37/75 primary bronchopulmonary tumours (49%) were identified as HPV positive by the PCR method: 18 cases were purely HPV 16 positive (49%), 12 were purely HPV 6 positive (32%), 5 cases were HPV 16/6 positive (14%), 1 case was HPV 16/11 positive (2%) and 1 case was HPV 16/18 positive (2%). Fourteen metastases were HPV positive, and HPV 16, 11 and 6 were detected in both regional and distant metastases. Two of the HPV 16-positive metastases were brain metastases from two separate HPV 16-positive primary tumours; 35% of the HPV-positive cases were adenocarcinomas, 30% squamous cell carcinomas, 22% oat cell carcinomas, 5% large cell carcinomas, 3% anaplastic carcinoma, 3% low-differentiated carcinoma, and 3% malignant cylindroma. The CIN III lesions from 34 of the 37 HPV-positive bronchopulmonary carcinomas were analysed by PCR. The overall HPV positivity in the CIN III lesions was 74% (25/34 cases): 48% were purely HPV 16 positive, 24% purely HPV 6 positive, 24% HPV 16/6 positive and 4% were HPV 18 positive. Our results indicate that HPV is also involved in the development of bronchopulmonary cancers in women with a history of CIN III lesions.     

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.     

Detection and genotype analysis of human papillomavirus in non-small cell lung cancer patients

Although the role of human papillomavirus (HPV) in the development of uterine cervical cancer is well established, the role of HPV in lung carcinogenesis remains controversial. The detection rates of HPV DNA are subject to a wide variation from 0 to 100 %. This is partly influenced by the detection techniques employed. To elucidate the impact of HPV infection on lung parenchyma, we analyzed 100 non-small cell lung cancer (NSCLC) specimens (39 squamous cell carcinomas, 50 adenocarcinomas, 5 samples with characteristics of both squamous cell and adenocarcinoma, 5 undifferentiated and 1 large cell carcinoma) from the region of Crete, Greece. Sixteen non-cancerous samples served as the negative controls. DNA was extracted from 100 paraffin-embedded tissue sections obtained from NSCLC patients. The specimens were examined for the detection of HPV DNA by Real-Time PCR using GP5+/GP6+ primers. Furthermore, the HPV-positive samples were subjected to genotyping. In contrast to the absence of viral genomes in the control samples, HPV DNA was detected in 19 NSCLC specimens (19 %). In particular, 4 squamous cell carcinomas, 12 adenocarcinomas, 1 sample with characteristics of both squamous cell and adenocarcinoma, and 2 undifferentiated samples were HPV-positive. The distribution of HPV genotypes was as follows: HPV 16: eight cases (42.1 %); HPV 11: three cases (15.8 %); HPV 6: one case (5.2 %); HPV 59: one case (5.2 %); HPV 33: two cases (10.5 %); HPV 31: two cases (10.5 %) and HPV 18: two cases (10.5 %). The presence of HPV in the tumor samples provides evidence of the potential role of HPV in NSCLC and strongly argues for additional research on this issue.     

Joint effects of different human papillomaviruses and Chlamydia trachomatis infections on risk of squamous cell carcinoma of the cervix uteri

This case-control study based in Nordic serum banks evaluated the joint effects of infections with genital human papillomavirus (HPV) types, and Chlamydia trachomatis in the aetiology of cervical squamous cell carcinoma. Through a linkage with the cancer registries, 144 cases were identified and 420 controls matched to them. Exposure to past infections was defined by the presence of specific IgG antibodies. The odds ratio (OR) for the second-order interaction of HPV16, HPV6/11 and C. trachomatis was small (1.0) compared to the expected multiplicative OR, 57, and the additive OR, 11. The interactions were not materially different among HPV16 DNA-positive squamous cell carcinomas. When HPV16 was replaced with HPV18/33 in the analysis of second-order interactions with HPV6/11 and C. trachomatis, there was no evidence of interaction, the joint effect being close to the expected additive OR. Possible explanations for the observed antagonism include misclassification, selection bias or a true biological phenomenon with HPV6/11 and C. trachomatis exposures antagonizing the carcinogenic effects of HPV16.     

Presence of Human Papillomavirus Type 18 DNA in a Pharyngeal and a Laryngeal Carcinoma

We examined the presence of human papillomavirus (HPV) genome in pharyngeal and laryngeal squamous cell carcinomas by employing the polymerase chain reaction. We detected HPV 16 DNA in one of 11 pharyngeal carcinomas and in 3 of 28 laryngeal carcinomas, and HPV 18 DNA in a pharyngeal and a laryngeal carcinoma. In one of the laryngeal carcinomas, DNA of both HPV types 16 and 18 were detected. To our knowledge, this is the first report of detection of HPV type 18 in head and neck carcinomas.     

Presence of human papillomavirus type 18 DNA in a pharyngeal and a laryngeal carcinoma.

We examined the presence of human papillomavirus (HPV) genome in pharyngeal and laryngeal squamous cell carcinomas by employing the polymerase chain reaction. We detected HPV 16 DNA in one of 11 pharyngeal carcinomas and in 3 of 28 laryngeal carcinomas, and HPV 18 DNA in a pharyngeal and a laryngeal carcinoma. In one of the laryngeal carcinomas, DNA of both HPV types 16 and 18 were detected. To our knowledge, this is the first report of detection of HPV type 18 in head and neck carcinomas.     

Increasing incidence of CD44v7/8 epitope expression during uterine cervical carcinogenesis

Splice variants of the cell surface glycoprotein CD44 (CD44v) have been implicated in the progression of various human tumors. In the present study, we have examined the expression pattern of a CD44v epitope encoded by the adjacent variant exons v7 and v8 during human cervical carcinogenesis. While only 1/11 normal cervical squamous epithelia was positive for this epitope by immunohistochemistry, 4/21 samples of low-grade squamous intra-epithelial lesions (LSIL), 17/35 samples of high-grade squamous intra-epithelial lesions (HSIL), 11/12 samples of the HSIL subgroup of carcinoma in situ and 17/17 cases of invasive cervical carcinoma showed CD44v7/8 epitope expression. In addition to CD44 variant expression, we have analyzed 67 lesions for the presence of HPV16/18-DNA using PCR. Most of the samples expressing the v7/8 epitope were also HPV16-positive (29/32), whereas only 17/35 of the v7/8-negative samples were HPV16-positive. HPV18 DNA was found in only one invasive carcinoma. Our data suggest that high-risk HPV infection may precede CD44v7/8 expression and that the number of samples expressing the CD44v7/8 epitope increases during carcinogenesis and reaches nearly 100% at the carcinoma in situ stage. This CD44 epitope could, therefore, serve as a diagnostic marker of cervical squamous cell carcinomas and as a possible target for CD44v7/8 epitope-directed therapies. © 1996 Wiley-Liss, Inc.     

Relationship between p21 and p53 expression, human papilloma virus infection and malignant transformation in sinonasal-inverted papilloma

AimsTo identify the relationship between p21 and p53 expression, human papilloma virus (HPV) infection and malignant transformation in sinonasal-inverted papilloma.Material and methodsNasal tissues, exophytic papilloma, inverted papilloma (IP) with dysplasia, IP with carcinoma and invasive squamous cell carcinoma (SCC) were stained with the monoclonal antibodies p21 and p53. In-situ hybridisation for HPV DNA was also carried out for types 6/11, 16/18 and 31/33.ResultsSignificant increased staining of p21 and p53 was observed in IP with severe dysplasia, IP with carcinoma and invasive carcinoma compared with control nasal mucosa. A significant increase of dysplasia was observed in IP in the HPV 6/11 and 16/18-positive group, compared with the HPV 6/11 and 16/18-negative group. Significant decrease in expression of p21 and p53 was observed in HPV 16/18-positive IP compared with HPV 16/18-negative IP.ConclusionsOur data raise the possibility that testing for p21, p53 and HPV may help to screen out papilloma lesions with a potential for dysplasia or carcinoma.