Human papillomavirus 16 in breast cancer of women treated for high grade cervical intraepithelial neoplasia (CIN III).

Women with both a history of high grade cervical intraepithelial neoplasia (CIN III) and breast carcinoma as second primary cancer were selected for studying the presence of HPV in breast carcinomas. Paraffin embedded material from 38 patients with 41 breast carcinoma cases after CIN III were examined by polymerase chain reaction (PCR) and in situ hybridization. By PCR we detected HPV 16 DNA in 19 out of 41 cases (46%) of the breast carcinomas. One case proved to be HPV 16 positive also by in situ hybridization. HPV 16 was also detected in 32 out of the 38 patients with CIN III (84%). All HPV 16 positive breast carcinomas were HPV 16 positive in their corresponding CIN III lesions. Eight patients with diagnosed breast cancer before the CIN III lesions were used as controls. None of these had HPV positive breast carcinomas. No cases were positive for HPV 11, 18, or 33. HPV 16 was detected in the primary tumours, in local metastases from HPV 16 positive tumours, in a distant HPV 16 positive breast carcinoma metastasis to the colon, and in other primary cancers in patients with HPV 16 positive breast carcinomas and HPV 16 positive CIN III. Estrogen and progesterone receptors were quantified in the HPV positive and HPV negative breast carcinomas, and there was no significant difference in the fraction positive in the two groups. Oncogenic HPV DNA might be transported from an original site of infection to other organs by blood or lymph, and possibly be a factor in the development of cancer in different organs.     

HPV 16 in multiple neoplastic lesions in women with CIN III

Paraffin embedded material of multiple primary cancers and other hyperplastic tumours from fifteen patients were analyzed by PCR and in situ hybridization for the presence of HPV DNA in the lesions. All patients had also high grade cervical intraepithelial dysplasia (CIN III) and breast carcinomas and were selected from a previous study enrolling 46 women with CIN III and breast carcinomas. HPV 16 was detected by PCR in 8/15 patients (53%), with eleven HPV 16 positive tumours. HPV 16 was detected in two malignant melanomas, one basal cell carcinoma, one squamous cell carcinoma of the vulva, one Bowen disease of the vulva, two high grade vaginal intraepithelial neoplasias, one cancer corporis uteri, one bronchial carcinoma and two lymphomas. Three cases, two high grade vaginal intraepithelial neoplasia and a squamous cell carcinoma of the vulva, were also reported to be positive by in situ hybridization. 5/8 patients (63%) with HPV 16 positive second cancers had also HPV 16 positive breast carcinomas. All fifteen patients with second cancers after CIN III had HPV 16 positive CIN III lesions; 53% of the patients had also a familial cancer history. We assume that HPV 16 may be involved in the development of different second cancers in women with HPV 16 positive CIN III.     

Coexisting high-grade glandular and squamous cervical lesions and human papillomavirus infections

The frequency of high-risk human papillomavirus (hr-HPV) genotypes in patients with adenocarcinoma in situ (ACIS) with coexisting cervical intraepithelial neoplasia (CIN), ACIS without coexisting CIN, and high-grade CIN (CIN II/III) was studied, in order to gain more insight into the relation between hr-HPV infections and the development of coexisting squamous and glandular lesions. The SPF(10) LiPA PCR was used to detect simultaneously 25 different HPV genotypes in biopsies obtained from 90 patients with CIN II/III, 47 patients with ACIS without coexisting CIN, and 49 patients with ACIS and coexisting CIN. hr-HPV was detected in 84 patients (93%) with CIN II/III, 38 patients (81%) with ACIS without CIN, and in 47 patients (96%) with ACIS and coexisting CIN. A total of 13 different hr-HPV genotypes were detected in patients with CIN II/III, and only five in patients with ACIS with/without coexisting CIN. HPV 31, multiple hr-HPV genotypes, and HPV genotypes other than 16, 18, and 45 were significantly more often detected in patients with CIN II/III, while HPV 18 was significantly more often detected in patients with ACIS with/without CIN. There were no significant differences in the frequency of specific hr-HPV genotypes between patients with ACIS with or without coexisting CIN. In conclusion, the frequency of specific hr-HPV genotypes is similar for patients with ACIS without CIN and patients with ACIS and coexisting CIN, but is significantly different for patients with CIN II/III without ACIS. These findings suggest that squamous lesions, coexisting with high-grade glandular lesions, are aetiologically different from squamous lesions without coexisting glandular lesions.     

HPV in situ hybridization: impact of different protocols on the detection of integrated HPV

Although there is consensus that HPV integration is common in invasive cervical carcinomas and uncommon or absent in low-grade uterine cervical intraepithelial neoplasia (CIN I), estimates for HPV integration in CIN II/III range from 5 to 100% using different PCR-based and in situ hybridization (ISH) approaches. It has been suggested that HPV integration can be identified using ISH by scoring of punctate signals. The increased sensitivity of fluorescence ISH (FISH) methods, allowing the detection of single copies of HPV, complicates the distinction between integrated and episomal HPV. Recently it has been suggested that, in such assays, the signals originating from integrated virus can be hidden in a background of episomal HPV. We therefore compared 2 different FISH protocols for the detection of integrated HPV in a series of CIN II/III lesions: 1) a mild protocol in which episomal HPV and RNA is retained and 2) a harsh protocol that extensively extracts proteins and RNA, and which promotes the partial loss of episomal HPV but not integrated HPV. A series of 28 HPV 16/18 positive CIN II/III lesions (17 solitary lesions and 11 lesions adjacent to microinvasive carcinoma) were studied. A punctate signal pattern was identified in 7 of these lesions with both protocols. Punctate signal was also present in control samples from lesions that are known to be associated with HPV integration (invasive squamous cell carcinoma (n = 3), adenocarcinoma in situ (n = 3), and invasive adenocarcinoma (n = 1). HPV RNA contributed significantly to the intensity of punctate FISH signal, especially when applying the mild protocol, as shown by omitting DNA denaturation, including RNase pretreatment steps and measuring the fluorescence signal intensity. Also, HPV RNA was frequently detected in addition to episomal/integrated HPV DNA in the majority of the other 21 CIN II/III lesions; this resulted in intense granular/diffuse FISH signals throughout the epithelium. However, in 7 of these lesions, the harsh protocol gave a more consistent punctate pattern in cells throughout the full thickness of the epithelium. This supports the hypothesis that the harsh protocol unmasks integrated HPV more efficiently by extracting RNA and episomal HPV. Overall, with this harsh protocol, a clonally expanded population of cells containing punctate HPV signals was found in 5 of 17 (29%) solitary CIN II/III lesions and in 9 of 11 (88%) CIN II/III lesions associated with microinvasive carcinoma. Combining these data with the results from our previous study, with the harsh protocol in 7 of 40 (18%) solitary CIN II/III lesions and 19/21 (90%) CIN II/III lesions associated with microinvasive carcinoma (p < 0.001), this pattern was found. This indicates that, when robustly defined, a punctate HPV pattern in CIN II/III lesions is associated with the presence of an invasive carcinoma.     

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.     

Assessment of cytotoxic T-lymphocyte phenotype using the specific markers granzyme B and TIA-1 in cervical neoplastic lesions.

Cervical carcinomas are closely associated with high-risk human papillomavirus (HPV) types and are preceded by cervical intraepithelial neoplasia (CIN). Most CIN lesions regress spontaneously and will not evolve to invasive carcinoma. The cellular immune system mediated by cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells are thought to play an important role in the ultimate decline of CIN lesions. Although TIA-1 is constitutively expressed in the majority of circulating T cells and defines a subpopulation of CD8+ T cells with cytotoxic potential, granzyme B is only expressed in CTLs upon activation. In the present study we have evaluated the expression of these proteins by lymphocytes present in 24 randomly chosen CIN lesions with increasing degree of atypia and in 14 cervical squamous cell carcinomas. As major histocompatibility complex (MHC) class I expression is frequently down-regulated in HPV-induced lesions, thus possibly frustrating tumour cell recognition by infiltrating CTLs, these lesions were also analysed for MHC class I expression. The results indicated that in most CIN lesions only a minority of CTLs are activated, whereas in some carcinomas a massive infiltration of activated, i.e. granzyme B-positive, CTLs were observed. The percentage of activated CTLs was not related to expression of MHC class I on neoplastic cells. These results suggest that in some carcinomas proper activation of CTLs occurs but that most likely local factors or immunoselection of resistant neoplastic cells inhibit a proper response of CTLs to these neoplastic cells.     

p16INK4a Immunoprofiles of Squamous Lesions of the Uterine Cervix–Implications for the Reclassification of Atypical Immature Squamous Metaplasia

p16^INK4a immunoprofiles of non-precancerous and dysplastic squamous cervical lesions were defined and applied to the reclassification of atypical immature squamous metaplasia (AIM). The immunoexpression of cytokeratin 17 (CK 17) in AIM was also evaluated. Totally, 295 cervical cone biopsies representing squamous metaplasia, reactive changes, koilocytosis, flat condyloma, CIN I, CIN II, CIN III and AIM were subjected to p16^INK4a immunohistochemistry. AIM cases were analyzed using CK 17 antibody. Typical p16^INK4a immunoprofiles for the metaplastic, LSIL/HPV and HSIL phenotypes were recorded and used for the categorization of AIM into particular phenotype groups. Results were correlated with CK 17 immunoexpression. All CIN II and CIN III lesions, all but one case of CIN I and all flat condylomas overexpressed p16^INK4a. Other non-precancerous lesions, including koilocytosis, were predominantly negative. Contrary to the sporadic and focal immunostaining, diffuse positivity was associated with the dysplastic features of the lesion. CIN II and CIN III were characterized by a diffuse, strong/weak, full-thickness staining, whereas CIN I showed a heterogeneous diffuse/focal, weak/strong, lower half positivity. One third of AIM lesions may be reclassified as HSIL, one third as LSIL/HPV and one third shows metaplastic phenotype. All AIM cases with metaplastic and LSIL/HPV phenotypes expressed CK 17 diffusely, whereas focal positivity slightly prevailed in AIM with HSIL phenotype. We conclude that p16^INK4a immunohistochemistry is a supporting method for the differential diagnosis of cervical lesions, which may be especially useful for the reclassification of AIM. The efficacy of CK 17 immunohistochemistry seems to be controversial for these purposes.     

Human papillomavirus 16 load in normal and abnormal cervical scrapes: an indicator of CIN II/III and viral clearance

The relation between human papillomavirus type 16 (HPV 16) viral load in cervical scrapes and development of high-grade cervical intraepithelial neoplasia (CIN II or III) was studied in a nested case-control study of women with normal cytology (group A) and in a cohort of women with abnormal cytology (group B). HPV 16 DNA load was determined using a quantitative real-time PCR assay. In group A, case women (women with CIN II/III, n = 12) had a significantly higher viral load than control women (women with CIN < or = I, n = 47). This resulted in an increased relative risk of women with the 50% highest viral load for development of CIN II/III (OR 7.7; CI 1.6-33). In group B, women with CIN II/III (n = 38) had a significantly higher viral load than women with CIN < or = I (n = 25). Women with the 50% highest viral load had an increased relative risk of CIN II/III (OR 3.2; CI 1.1-9.3) and a decreased chance of both viral clearance and cytologic regression. Our data suggest that in women with normal cytology an increased HPV 16 load confers an increased risk of developing a CIN lesion. A sustained high viral load is subsequently informative for progression to a high-grade CIN lesion.     

Detection of human papillomavirus types 6/11, 16 and 18 in exfoliated cells from the uterine cervices of Japanese women with and without lesions

Human papillomavirus (HPV) infection of the uterine cervices of Japanese women with and without lesions was identified by the filter in situ hybridization method. Exfoliated cervical cells from 23 cervical papillary condylomas, 70 cervical intraepithelial neoplasia (CIN) grade I/II, 26 CIN III, 31 invasive cervical cancers and 666 cervices without evidence of disease (including 53 pregnant women) were tested for the presence of HPV types 6/11, 16 and 18. The positive rates for the detection of HPV types 6/11, 16 and 18 DNA were 47.8%, 26.1% and 8.7% in cervical condylomas, 5.7%, 15.7% and 8.6% in CIN I/II, 0, 34.6% and 0 in CIN III, 3.2%, 38.7% and 9.7% in invasive cervical cancers and 0.9%, 1.8% and 0.6% in the control cervices. These data suggest that, in Japan, HPV6/11, HPV16 and HPV18 infections are also prevalent in cervical cells with normal phenotype, and the type of HPV infection of the uterine cervix is related to the histological diagnosis.     

Detection of Human Papillomavirus Types 6/11, 16 and 18 in Exfoliated Cells from the Uterine Cervices of Japanese Women with and without Lesions

Human papillomavirus (HPV) infection of the uterine cervices of Japanese women with and without lesions was identified by the filter in situ hybridization method. Exfoliated cervical cells from 23 cervical papillary condylomas, 70 cervical intraepithelial neoplasia (CIN) grade I/II, 26 CIN III, 31 invasive cervical cancers and 666 cervices without evidence of disease (including 53 pregnant women) were tested for the presence of HPV types 6/11, 16 and 18. The positive rates for the detection of HPV types 6/11, 16 and 18 DNA were 47.8%, 26.1% and 8.7% in cervical condylomas, 5.7%, 15.7% and 8.6% in CIN I/II, 0, 34.6% and 0 in CIN III, 3.2%, 38.7% and 9.7% in invasive cervical cancers and 0.9%, 1.8% and 0.6% in the control cervices. These data suggest that, in Japan, HPV6/11, HPV16 and HPV18 infections are also prevalent in cervical cells with normal phenotype, and the type of HPV infection of the uterine cervix is related to the histological diagnosis.     

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.     

Prevalence of Human Papillomavirus in Abnormal Cervical Smears in Malaysian Patients

Cervical cancer is the second most common female malignancy in Malaysia. Despite advances in treatment,the overall survival for this disease has not changed in the last decade. Infection by certain types of HPV isrecognized as a causal and necessary factor for its development. This study was carried out to determine theprevalence of HPV infection in abnormal cervical smears in Malaysian patients using archival cervical smearsretrieved from the Cytopathology Unit, Universiti Kebangsaan Malaysia Medical Centre (UKMMC) betweenthe years 1992-1995. DNA was extracted from 38 abnormal smears comprising 25 intraepithelial lesions and 13cervical carcinomas and 10 normal smears. Amplification of HPV genes was carried out using the polymerasechain reaction (PCR) technique. HPV genotypes were determined using direct sequencing and the results werecompared to the database from Genebank. DNA was successfully extracted from all 48 cervical smears. HighriskHPV (HR-HPV) genotypes were detected in 95% of the abnormal smears. Eight high-risk oncogenic typeswere identified: 16, 18, 31, 51, 52, 56, 58 and 66. All (100%) cervical cancer smears showed presence of HR-HPVcompared to 92% of the cervical intraepithelial lesions. Among the eight HR-HPV genotypes identified, HPV 16and 52 were the commonest (23.7% each) HPV genotypes encountered and among the CIN lesions, HPV 16(28%) was the most frequent. We conclude that HPV 16 is the most prevalent HPV genotype present in abnormalcervical smears in Malaysian patients, and that the use of archival material to assess the presence of HPV ispotentially worthwhile, and can be utilized for longitudinal studies of HPV presence and persistence.     

Human papillomavirus DNA in cervical intraepithelial neoplasia detected by in situ hybridisation

Human papillomavirus (HPV) infection was investigated by in situ hybridisation in histological sections from 38 women with abnormal Papanicolaou smears. 13 patients had condylomatous lesions without atypia, 15 cervical intraepithelial neoplasia (CIN) I, 4 CIN II, 3 CIN III and 2 carcinoma in situ (CIS). HPV DNA was detected in 29 cases (78%) (1 specimen was technically inadequate). HPV 16 and 18, and 31, 33 and 35 were both present (67%) in CIN III. HPV 6 and 11 were more frequent in CIN I (56%) and in condylomatous lesions (38%). 31% of the condylomatous lesions without atypia contained HPV 31, 33, and 35 and 31% of those with CIN I were infected with HPV 16 and 18. These data confirm the frequent association of HPV infection with cervical cancer and CIN, and indicate that in situ hybridisation can identify patients with specific types of HPV infection at risk for cervical cancer.     

Risk Categorization with Different Grades of Cervical Pre-Neoplastic Lesions - High Risk HPV Associations and Expression of p53 and RARβ

Objective: To identify high risk HPV associations by evaluating linked p16 overexpression and also the expressionof p53 and RARβ together with histopathology for risk categorization of cervical pre-neoplastic lesions. Materials andMethods: Immunohistochemical staining was performed on 100 cases of cervical pre- neoplastic lesions for expressionof biomarkers like p16, p53 and RARβ for comparison with haematoxylin/eosin (HE) findings. All the experimentallygenerated data were statistically analyzed. Results: In this study 70% cases showed overexpression of p16INK4Aincreasing progressively from CIN I to CIN II but reduced in CIN III (p <0.01). p53 oncoprotein expression was seenin 51% cases, again with increments from CIN I to CIN II with slight reduction in CIN III (p<0.01). Some 24% casesshowed negative immunoreactivity for the putative tumor suppressor gene RARβ (p>0.05). Conclusion: Our studyprovides support for the idea that p16 can be used to identify associations with HPV , as well as having potentialalong with p53 and RARβ for categorizing cervical pre-neoplastic cases having a higher risk of neoplastic conversion.Thus it may be concluded that accurate risk categorization can be achieved with the help of genetic markers as wellas histopathology.     

A review of the natural history of cervical intraepithelial neoplasia

Cervical Intraepithelial Neoplasia (CIN) is a premalignant lesion of cervical squamous cell carcinoma which over time may persist unchanged, regress to normal or a lesser grade of CIN, or progress to a higher grade of CIN or invasive carcinoma. Rates of progression correlate directly with the CIN grade. Human Papilloma Virus (HPV) detection is a significant determinant of CIN regardless of grade. Studies of risk factor profiles, cytogenetic abnormalities, cell proliferation indices, cell cycle and senescence control, oncogene and tumor suppressor gene expression, protein expression, and HPV status have been conducted to identify determinants of CIN I and CIN II/III and predictors of CIN 1 progression. Differences in these attributes suggest that CIN I is a sexually transmitted, productive HPV infection, whereas CIN II/III is a dysplastic lesion resulting from repeated exposure to a sexually transmitted HPV and possibly an additional agent. HPV16 positivity and increased viral load in some earlier studies were predictive of prevalent CIN II/III. More recent studies with more sensitive HPV assays did not corroborate these findings. The role of cigarette smoking is controversial and requires additional study. Accumulating evidence suggests that high risk HPV DNA detection and persistence are predictive of CIN I progressing to CIN II/III. Other possibilities are persistence of a high risk HPV variant, altered cell immunity, and cigarette and oral contraceptive use. Possible biomarkers include aneuploidy, aneusomy of chromosomes 1 and 3, Ras and bcl-2 oncogene over expression, and cytokeratin 13 protein under-expression.     

Cytologic detection of condylomas and cervical intraepithelial neoplasia of the uterine cervix with histologic correlation

Human papillomavirus (HPV) has been implicated as an important etiologic factor in cervical carcinoma. This study evaluates the efficacy of cytology as a screening tool in the detection of cervical lesions with koilocytotic features. Cervical smears and biopsy specimens from 76 women seen between January 1983 and October 1985 were reviewed. The histologic categories consisted of koilocytotic lesions (flat condylomas) with minimal cellular atypia, CIN I, II, III, with surface koilocytes showing cellular atypia (atypical koilocytosis), CIN III with a contiguous lesion as defined in categories 1 and 2, negative biopsies. Histologically, five cases showed flat condylomas with minimal cellular atypia, 65 showed cervical intraepithelial neoplasia (CIN) with atypical koilocytosis and two showed negative biopsies. Cytologically, in all cases of flat condyloma with minimal cellular atypia (5+5), CIN I with atypical koilocytosis (39/39), and CIN III with a contiguous condylomatous lesion (4/4) both koilocytes and, in the latter two categories, dysplastic cells were identified in the cervical smears. In 7/9 cases of CIN II and 9/17 cases of CIN III with atypical koilocytosis, smears showed both atypical koilocytes and dysplastic cells. In the remaining ten cases, however, there was cytologic underestimation of the histologic diagnosis of CIN, particularly when the lesion was focal. The data suggests that although cytology does detect a high percentage of cervical lesions with koilocytotic features (64/76, 84% in this study), it may not detect focal CIN II, CIN III lesions associated with condylomatous lesions in the same biopsy specimen. Therefore, it is proposed that all women with cytologic evidence of koilocytosis on cervical smears are deserving of a colposcopic examination.     

Type distribution, viral load and integration status of high-risk human papillomaviruses in pre-stages of cervical cancer (CIN)

A series of 176 archival cervical intraepithelial neoplasia (CIN) was analysed for the presence, viral load and integration status of 'high-risk' types of human papillomavirus (HR-HPV). The samples were assayed using newly developed methods based on real-time PCR. Two methods for the extraction of DNA from the paraffin-embedded biopsies were compared: a protocol based on the MagNA pure system (Roche) and a Qiagen spin column kit (Qiagen). It was possible to amplify 94% (166) of the samples. Of these, 36, 63 and 80% of the CIN I, II and III cases contained HR-HPV. HPV 16 was the most prevalent, and was found in 20, 28 and 46% of the CIN I, II and III cases, respectively. The second most frequent HR-HPV was type 33 group, and in CIN II it was as prevalent as HPV 16. The median number of copies of HR-HPV per cell was not significantly different in the CIN I, II and III cases, but there was a wide range of viral load values over several magnitudes, regardless of the grade of CIN. All samples were found to contain integrated forms of HPV 16, frequently mixed with an episomal form.     

血管内皮生长因子-C、高危型人乳头瘤病毒在子宫颈上皮内瘤样病变及子宫颈鳞癌中的表达和意义

 目的 探讨人类乳头状瘤病毒（HPV）感染和血管内皮生长因子-C（VEGF-C）在宫颈癌的形成及演进过程中可能的作用机制。方法 采用PCR技术检测16例正常宫颈，65例宫颈上皮内瘤样病变（CIN），其中，20例CINI组、24例CINⅡ组、21例CINⅢ组，26例宫颈浸润性癌（宫颈癌组）组织中HR-HPV的感染情况，同时采用免疫组化SABC法检测VEGF-C的表达水平。结果 VEGF-C在CIN在及宫颈鳞癌组织中均表达，并且随着病变程度的增强VEGF-C的表达呈渐进性增强，而正常的宫颈组织细胞中无表达。HR-HPV同CIN及宫颈癌均显著相关（比值比分别为19.12和20.49；95 %CI分别为2.31 ~ 157.8和3.28 ~ 226.09）。CIN及宫颈癌组织中VEGF-C的表达同HR-HPV的感染显著相关。结论 VEGF-C蛋白的表达同HR-HPV感染显著相关，可能是宫颈癌血管形成的早期预测指标。