Prevalence and distribution of human papillomavirus type-16 DNA in pelvic lymph nodes of patients with cervical cancer and in women with no history of cervical abnormality.

The polymerase chain reaction (PCR) was used to investigate the prevalence and distribution of human papillomavirus (HPV)-16 DNA in paraffin sections of all pelvic lymph nodes removed from 14 patients with Stage Ib-cervical cancer at the time of resection of their primary tumours. The results were compared with those obtained from 8 women with no known history of cervical abnormality. In all, 22 cervical biopsies and 40 I lymph nodes (296 paraffin blocks) were examined. Nine of the 14 cervical cancer patients had primary tumours that were positive for HPV-16 DNA: only 3 of these had lymph nodes with histological evidence of metastasis, and HPV 16 DNA was detected in each of the corresponding paraffin blocks. HPV 16 DNA was also detected in varying proportions (8%-92%) of the histologically-negative lymph nodes from these women. There was no correlation between the HPV DNA-positive lymph nodes and their proximity to the primary tumour. HPV-16 DNA was not identified in any of the lymph nodes from the 5 women whose cancers were not HPV-16-related, or in those of women with no evidence of cervical abnormality. This preliminary survey suggests that HPV DNA is frequently transported from HPV-16-related cervical tumours to regional lymph nodes. However, its practical significance will not be clear until sufficient time has elapsed for correlation of the results with the clinical outcome.     

Correlation between Load of HPV 16 DNA in Cervical Cancer and HPV 16 DNA in Lymph Nodes

OBJECTIVE To determine the association between viral load of human papillomavirus 16 (HPV16) DNA in the primary focus of cervical carcinoma and HPV16 DNA in pelvic lymph nodes. METHODS The HPV16 DNA load was measured by fluorescent quantisation polymerase chain reaction (FQ-PCR) in 17 primary foci. HPV16 DNA was detected by polymerase chain reaction (PCR) using HPV16 type-specific primers in 296 pelvic lymph nodes which were from 17 cases of cervical cancer. RESULTS The viral load of HPV16 DNA showed statistically significant differences between tumors with a diameter of < 4 cm and > 4 cm (P < 0.05). Seven of 17 cervical cancer cases had HPV16 DNA positive lymph nodes, designated as the positive group, while the remaining 10 without positive lymph nodes was designated the negative group. The average load of HPV16 DNA showed no significant difference between the 2 groups (P > 0.05). The load of HPV16 in the primary lesion was not associated with that in the lymph nodes. There were 38 HPV16 DNA positive nodes in the total 296 nodes. The rate of positivity of HPV16 DNA in lymph nodes showed statistically significant differences in consideration of maximum tumor diameter, tumor differentiation, histologic type, depth of myometial infiltration and the metastatic status of the nodes, respectively (P < 0.05).CONCLUSION Viral load of HPV16 in the primary cancer focus correlated with the quantity of tumor cells in the primary focus but not with the existence of HPV DNA positive lymph nodes. Detection of HPV DNA may help to find the early metastases that cannot be evaluated histopathologically but the prognostic value of HPV positive lymph nodes needs further examination.     

子宫颈癌患者血清、盆腔淋巴结HPV检测及其相关性探讨

目的：检测宫颈癌患者原发灶、血清及其盆腔淋巴结中HPVDNA及亚型,探讨其相关性及临床意义。方法：选取16例行广泛全子宫切除术和盆腔淋巴结清扫术宫颈癌患者的原发灶组织、术前静脉血与盆腔淋巴结石蜡组织,运用PCR方法对上述标本进行HPVDNA及亚型的检测。结果：宫颈癌原发灶组织、血清标本中HPVDNA阳性率为50％（8／16）。16例盆腔淋巴结石蜡组织中13例为HPVDNA阳性（13／16,81．25％）,其中总共切除的133个淋巴结中60个为阳性（60／133,45．1％）,8例淋巴结HPVDNA阳性的病例其对应的原发灶组织也为阳性且两者亚型相同。盆腔淋巴结中的HPVDNA阳性率为45．1％（60／133）,显著高于病理证实的淋巴结转移率1．5％（2／133）。6例患者（6／16,37．5％）原发灶、血清、盆腔淋巴结同时均为HPVDNA阳性;2例患者（2／16,12．5％）原发灶、盆腔淋巴结中HPVDNA表达阳性,而血清为阴性;1例患者（1／16,6．25％）淋巴结、血清HPVDNA阳性,而原发灶为阴性;未发现原发灶、血清HPVDNA表达阳性而淋巴结为阴性的病例,而且以上HPVDNA阳性的病例同一个患者对应的HPV亚型也相同。结论：宫颈癌患者血清中HPVDNA检出率与临床分期无关。宫颈癌患者盆腔淋巴结的HPVDNA检测可提高病理诊断淋巴结转移的阳性率,并且淋巴结中HPVDNA的检出率与原发灶的分化程度相关。宫颈癌原发灶、血清、盆腔淋巴结中HPV感染可能存在相关性。     

子宫颈癌患者血清、盆腔淋巴结HPV检测及其相关性探讨

目的:检测宫颈癌患者原发灶、血清及其盆腔淋巴结中HPV DNA及亚型,探讨其相关性及临床意义。方法:选取16例行广泛全子宫切除术和盆腔淋巴结清扫术宫颈癌患者的原发灶组织、术前静脉血与盆腔淋巴结石蜡组织,运用PCR方法对上述标本进行HPV DNA及亚型的检测。结果:宫颈癌原发灶组织、血清标本中HPV DNA阳性率为50%(8/16)。16例盆腔淋巴结石蜡组织中13例为HPV DNA阳性(13/16,81.25%),其中总共切除的133个淋巴结中60个为阳性(60/133,45.1%),8例淋巴结HPV DNA阳性的病例其对应的原发灶组织也为阳性且两者亚型相同。盆腔淋巴结中的HPV DNA阳性率为45.1%(60/133),显著高于病理证实的淋巴结转移率1.5%(2/133)。6例患者(6/16,37.5%)原发灶、血清、盆腔淋巴结同时均为HPV DNA阳性;2例患者(2/16,12.5%)原发灶、盆腔淋巴结中HPV DNA表达阳性,而血清为阴性;1例患者(1/16,6.25%)淋巴结、血清HPV DNA阳性,而原发灶为阴性;未发现原发灶、血清HPV DNA表达阳性而淋巴结为阴性的病例,而且以上HPV DNA阳性的病例同一个患者对应的HPV亚型也相同。结论:宫颈癌患者血清中HPV DNA检出率与临床分期无关。宫颈癌患者盆腔淋巴结的HPV DNA检测可提高病理诊断淋巴结转移的阳性率,并且淋巴结中HPV DNA的检出率与原发灶的分化程度相关。宫颈癌原发灶、血清、盆腔淋巴结中HPV感染可能存在相关性。     

Human papillomavirus 16 dna in cervical cancers and in lymph nodes of cervical cancer patients: A diagnostic marker for early metastases?

Human papillomavirus (HPV) 16 is most prevalent in cervical cancers and also persists in metastases. We examined HPV16-DNA-positive primary cancers and several lymph nodes from each of 14 patients to evaluate the use of HPV16 DNA as a diagnostic marker for the detection of early node involvement. The HPV16 DNA was exclusively integrated in 39% of the primary cancers, predominantly episomal in 36%, and integrated and extrachromosomal to a similar extent in 25%. Thirteen of 16 involved lymph nodes contained HPV16 sequences. Integrated viral DNA showed the same pattern in primary tumors and in metastases. The level of extrachromosomal HPV16 DNA, however, appeared to be considerably reduced in some nodes. HPV16 DNA was also detected in 18 out of 59 histologically negative lymph nodes. This result recommends nucleic acid hybridization as a sensitive method for the detection of HPV-DNA-positive cancer cells. The prognostic significance of viral sequences in histologically negative nodes remains to be established.     

早期宫颈癌前哨淋巴结中高危型HPV16/18 DNA表达的检测及临床意义

目的：探讨早期宫颈癌患者前哨淋巴结（SLN）中人乳头状瘤病毒（HPV）16/18 DNA表达检测对于微转移的临床意义。方法选取72例早期宫颈癌患者,予患者均行广泛性子宫切除加双侧盆腔淋巴结清扫术,术中采用染料法识别SLN的宫颈癌患者有46例,应用基因检测法（FQ-PCR）检测SLN中HPV16/18 DNA阳性表达情况,并分析其与各种临床病理因素的关系;对SLN病理阴性的33例患者进行长期随访,分析SLN中HPV16/18 DNA阳性与淋巴结转移的关系。结果46例宫颈癌患者SLN中HPV16/18 DNA阳性表达者共22例,其中13例淋巴结病理阳性患者中有10例阳性,而33例淋巴结病理阴性患者中仅12例阳性（P=0.013）;46例患者共检出前哨淋巴结102枚,均用FQ-PCR法检测HPV16/18 DNA,结果13例淋巴结病理阳性患者检出的37枚SLN中有29枚HPV16/18 DNA阳性,而33例淋巴结病理阴性患者检出的65枚SLN中仅有36枚阳性（P=0.033）;分析46例成功检出SLN的早期宫颈癌患者的临床资料,发现SLN中HPV16/18 DNA阳性表达仅与临床分期有关,具有统计学意义（P=0.034）;长期随访33例SLN病理阴性的患者,发现HPV16/18 DNA阳性的患者复发率高于HPV16/18 DNA阴性的患者,具有统计学意义（P=0.02）。结论检测宫颈癌SLN组织中HPV16/18 DNA表达可能是预测早期宫颈癌淋巴结微转移的可行方法。     

Prognostic value of human papilloma virus DNA test in patients surgically treated for squamous cell cervical cancer

The human papilloma virus (HPV) DNA test was performed in paraffinized biopsy samples of iliac lymph nodes obtained from 98 patients with cervical cancer receiving therapy in N. N. Petrov Research Institute of Oncology in 2000 to 2007. The data obtained was compared to patients clinical course. The HPV DNA was detected in 29 of 98 patients samples (29,6%), 27 of those patients had metastatic disease. These patients had significantly higher metastasis rate than HPV DNA-negative patients (p < or = 0,05). The following highly oncogenic genotypes of HPV DNA were detected: type 16 in 15 patients (51,72%), type 18 in 4 patients (13,79%), type 31 in 5 patients (17,24%) and type 33 in 5 patients (17,24%). In all the cases the HPV type detected in the lymph nodes corresponded to the one detected in the primary tumor. The relapse-free survival of HPV DNA-positive patients with iliac lymph nodes metastases (N1) was significantly lower than in HPV DNA-negative patients (p < or = 0,05). The iliac lymph nodes HPV DNA detection had 43,6% sensitivity (95% CI: 28/60%) and 79,7% specificity (95% CI: 67/89%) for relapse prediction. The test results had diagnostic value in 65,3% of cases, the test was false-negative in 56,4% and false-positive in 20,3% of cases.     

Ⅰa-Ⅱa期宫颈癌患者盆腔淋巴结中高危型HPVDNA及CK19的检测及其意义

目的：分别采用原位杂交法和免疫组化法检测早期宫颈癌盆腔淋巴结中高危型HPVDNA和CK19的表达,探讨早期宫颈癌微小转移的检测率及检测方法。方法：选取28例早期宫颈癌患者常规病理光镜检查证实转移和未转移淋巴结共104个,分别采用原位杂交法和免疫组化法检测高危型HPVDNA和CK19的表达。结果：所有常规病理检查阴性的80个淋巴结中高危型HPVDNA的阳性检出率为45％（36／80）;CK19的检出率为25％（20／80）。常规病理光镜检查证实淋巴未转移患者的57个淋巴结中23个高危型HPVDNA阳性（43．5％）,来自15名患者中的9人（60%）;14个淋巴结CK19阳性（24．6％）,来自15名患者中的7名（46．6％）。患者的微转移检出率分别为60％和46．6％。两种检测方法的结果有较好的一致性。高危型HPVDNA的检出率高于CK19（P〈0．001）。结论：原位杂交法检测高危型HPVDNA,免疫组化法测CK19均可检测出早期宫颈癌淋巴微转移．玛班涤交法是分子水平的检测,在宫颈癌淋巴微转移的检测中可能更敏感可靠。     

Relatively Low Prevalence of Human Papillomavirus 16, 18 and 33 DNA in the Normal Cervices of Japanese Women Shown by Polymerase Chain Reaction

Ninety-two cervical scrapes and tissues, obtained from cytologically or histologically normal cervices of Japanese women, were examined for the presence of human papillomavirus (HPV) 16, 18 and 33 DNA by the polymerase chain reaction (PCR) method. Five out of 92 cases were HPV 16 DNA-positive, but neither HPV 18 nor 33 DNA was detected. The HPV (type 16, 18 and 33) prevalence rate in pregnant women, including postpartum, was 10% (3/31), which was higher than that in non-pregnant women. In two HPV 16-positive cases, we detected HPV 16 DNA again 2 months later. HPV (type 16, 18 and 33) prevalence in normal cervices was shown to be relatively low. However, it is very important to follow up the HPV-positive cases in cytologically normal cervices in order to elucidate the relation between HPV infection and the progression of cervical cancer.     

Relatively low prevalence of human papillomavirus 16, 18 and 33 DNA in the normal cervices of Japanese women shown by polymerase chain reaction.

Ninety-two cervical scrapes and tissues, obtained from cytologically or histologically normal cervices of Japanese women, were examined for the presence of human papillomavirus (HPV) 16, 18 and 33 DNA by the polymerase chain reaction (PCR) method. Five out of 92 cases were HPV 16 DNA-positive, but neither HPV 18 nor 33 DNA was detected. The HPV (type 16, 18 and 33) prevalence rate in pregnant women, including postpartum, was 10% (3/31), which was higher than that in non-pregnant women. In two HPV 16-positive cases, we detected HPV 16 DNA again 2 months later. HPV (type 16, 18 and 33) prevalence in normal cervices was shown to be relatively low. However, it is very important to follow up the HPV-positive cases in cytologically normal cervices in order to elucidate the relation between HPV infection and the progression of cervical cancer.     

Early stage cervical cancers containing human papillomavirus type 18 DNA have more nodal metastasis and deeper stromal invasion.

PURPOSE: To investigate the clinical and pathological factors which might explain the poor prognosis associated with early stage cervical cancers containing human papillomavirus (HPV) type 18 DNA. EXPERIMENTAL DESIGN: A clinical and pathological review of 144 patients with stage IB cervical cancer treated with radical hysterectomy and bilateral pelvic lymph node dissection was done. HPV genotyping was determined from fresh tumor specimens through PCR. Clinical-pathological information, sites of recurrence, use of adjuvant radiation, and survival data were analyzed. RESULTS: Thirty-three (23%) tumors contained HPV 18 DNA. These tumors did not differ from those which contained non-HPV 18 DNA with respect to tumor grade or size. However, HPV 18-containing cancers were more likely to be adenocarcinomas. A higher incidence of pelvic lymph node metastasis was noted among the HPV 18 group (48%) as compared with the non-HPV 18 group (28%), and deeper stromal invasion was more common in HPV 18-associated tumors. Although a slightly higher proportion of patients with HPV 18-containing tumors received adjuvant radiation (67%) than those with non-HPV 18 cancers (49%), recurrences were more common among HPV 18 patients. Eleven (33%) of HPV 18-containing cancers relapsed compared with 18 (16%) of non-HPV18-containing tumors. CONCLUSIONS: The explanation for the worse prognosis associated with stage IB cervical cancers containing HPV 18 DNA treated with radical hysterectomy and bilateral pelvic lymph node dissection appears to be related to deeper cervical stromal invasion and more nodal metastases. Despite an increased use of adjuvant radiation therapy, these cancers are still more likely to relapse.     

Detection and quantitation of human papillomavirus DNA in the plasma of patients with cervical carcinoma.

Human papillomaviruses (HPVs) play a central role in the development of cervical carcinoma. Plasma DNA from 232 patients taken at diagnosis or after treatment for invasive cervical cancer (n = 175) or carcinoma in situ (n = 57) and 60 normal controls were examined for HPV-16 or HPV-18 E7 DNA by conventional and real-time quantitative PCR assays. We found HPV-16 or HPV-18 E7 DNA in 6.9% (11 of 175) of invasive cervical cancer cases (18.1% of cases positive for HPV-16 or HPV-18 at the genital tract), 1.8% (1 of 57) of carcinoma in situ, and 1.7% (1 of 60) of normal controls by conventional PCR. Quantitative PCR identified the highest concentrations of HPV DNA (copy number of HPV/ml of plasma) in patients with invasive cervical cancer (mean, 11,163; median, 183.5), followed by a level of 8 in the single carcinoma in situ case and 0 copies in the normal control initially positive by conventional PCR. HPV DNA can be detected in the plasma of some patients with HPV-positive cervical tumors. It remains to be demonstrated whether quantitative PCR analysis of HPV DNA in plasma may have utility in patients at high risk of recurrent disease.     

Frequency and prognostic significance of HPV DNA in sentinel lymph nodes of patients with cervical cancer.

BACKGROUND: It has been suggested that histologically undetectable or 'occult' metastases in the lymphatic system could explain some recurrences. HPV DNA screening by means of the polymerase chain reaction (PCR) has been proposed as a method to detect occult metastases. This study was designed to determine the frequency of HPV DNA detection by PCR in sentinel lymph node (SN), and its relation to the clinical characteristics and outcome of women with cervical cancer. PATIENTS AND METHODS: The primary cervical tumor and SN were tested for HPV DNA by means of PCR in 59 patients. RESULTS: Fifteen (25.4%) of the 59 women undergoing the SN procedure had an involved SN. HPV DNA was more frequent in positive SN than in negative SN (P < 0.0001). Seven patients had a recurrence, after a mean delay of 17 months (range: 10-26). One of seven patients with a recurrence had an involved SN. HPV DNA was detected in an SN of one of seven patients with recurrence and nine (19.5%) of 46 patients without recurrence (not significant). CONCLUSION: In women with cervical cancer, HPV DNA screening of sentinel nodes might help to identify patients at risk of lymph node metastases and recurrence.     

Expression of high-risk HPV DNA and CK19 in pelvic lymph nodes in stage Ia-IIa cervical cancer and their clinical value

The aim of this study was to investigate the detection rate and methods of micrometastases in early-stage cervical cancer by detecting the expression of high-risk HPV DNA and CK19 in pelvic lymph nodes. A total of 104 lymph nodes with/without pathologically confirmed metastases, from 28 patients with early-stage cervical cancer, were included for detection of high-risk HPV DNA and CK19 expression using in situ hybridization and immunohistochemistry, respectively. The detection rate of high-risk HPV DNA and CK19 in lymph nodes in patients with pathologically-confirmed lymph node metastases was higher compared to that in lymph nodes in patients without pathologically-confirmed lymph node metastases (P<0.001). In all 80 pathologically-negative lymph nodes, the positivity rates of high-risk HPV DNA and CK19 detection were 45 and 25%, respectively. In 57 lymph nodes in patients without pathologically-confirmed lymph node metastases the positivity rates of high-risk HPV DNA and CK19 detection were 43.5 and 24.6%. The detection rate of high-risk HPV DNA and CDK19 in 15 patients without pathologically-confirmed lymph node metastases were 60 and 46.6%, respectively. The detection rates of high-risk HPV DNA and CK19 in 104 lymph nodes were 56.7 and 41.3% (KI=0.46). The results of the two detection methods showed good consistency. Both detection of high-risk HPV DNA by in situ hybridization, and CK19 by immunohistochemical method detected lymph node micrometastases in early-stage cervical cancer. As a method of detection on the molecular level, in situ hybridization was more sensitive for the detection of lymph node micrometastases in early-stage cervical cancer.     

Relation between HPV-DNA and expression of p53, bcl-2, p21WAF-1, MIB-1, HER-2/neu and DNA ploidy in early cervical carcinoma: correlation with clinical outcome

The purpose of the present study was to analyze the relation between the expression of p53, bcl-2, p21WAF1, MIB-1, HER-2/neu, DNA ploidy and HPV16 or 18 infections with clinical parameters. HPV-DNA was evaluated in 171 early cervical carcinomas treated from 1965 to 1990 and detected by PCR (polymerase chain reaction) on paraffin specimens obtained before therapy was started. HPV-DNA of any type was detected in 78% (86/110) of all tumors, HPV16 was the predominant type and was seen in 56% (62/110), HPV18 in 8% (9/110) and HPV35 in 21% (23/110). Patients with HPV16 or 18 were significantly (P=0.011) younger than patients with tumors not containing these two HPV subtypes. Lymph node metastases were seen more frequently (P=0.047) in tumors expressing HPV16 or 18. Tumor size was associated with the HPV-type. The frequency of DNA aneuploidy was lower in high-risk HPV tumors than in tumors with other HPV subtypes (P=0.014). MIB-1 expression was highly significantly (P=0.00007) associated with presence of HPV16 or 18. The cancer-specific survival rate was lower for patients with HPV16 and 18 positive tumors, but the difference was not statistically significant. The overall 5-year survival rate of the complete series was 91%. In conclusion, the HPV DNA subtype was a prognostic factor in early stage cervical cancer and it was associated with age, positive lymph nodes, tumor size, DNA ploidy and the proliferation marker MIB-1.     

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.     

早期宫颈鳞癌癌周组织及盆腔淋巴结中podoplanin及高危型HPV的检测及意义

目的：检测早期宫颈鳞癌癌周组织及盆腔淋巴结中podoplanin及HPV16/18抗原的表达,寻求早期宫颈癌淋巴转移的标记物.方法：选取宫颈鳞癌57例,每例取1-3个淋巴结,共选取淋巴结103个,采用免疫组化方法检测podoplanin及HPV16/18的表达.结果：在有淋巴结转移的宫颈鳞癌癌周组织、无淋巴结转移的宫颈鳞癌癌周组织及正常宫颈组织中podoplanin的阳性率分别为76.2％（16/21）,41.7％ （15/36）,27.2％（6/22）.在淋巴转移患者的转移淋巴结,未转移淋巴结,及淋巴无转移患者的淋巴结中HPV16/18的阳性率分别为89.3％ （25/28）,60.0％ （18/30）,42.2％（19/45）,差异有统计学意义（P＜0.05）.二者的检测结果呈正相关.结论：联合检测盆腔淋巴结中HPV16/18和podoplanin抗原,有助于早期发现宫颈癌淋巴结微转移.Podoplanin可以作为检测宫颈癌淋巴转移的特异性标志.