Human papillomavirus DNA in invasive carcinoma of the vagina.

The presence of human papillomavirus (HPV) DNA in invasive carcinomas of the vagina, in their lymph node metastases, and in corresponding normal tissue was investigated by Southern blot hybridization with 32P-labeled HPV DNA. Tumor tissue from ten of 18 patients with vaginal carcinoma contained HPV DNA. Three of the 18 patients had a history of cervical neoplasia more than 14 years before the diagnosis of vaginal carcinoma. Five of 15 primary squamous cell carcinomas, one primary adenocarcinoma, and a vulvar recurrence of a vaginal squamous carcinoma contained HPV 16. A primary squamous carcinoma yielded HPV-related sequences. The HPV copy number varied from 0.5 to 50 per cellular genome. Four histologically positive inguinal lymph nodes from three patients contained HPV DNA. In six tumor-free control tissues from four patients, no HPV DNA was detected. No relationship was established between HPV positivity, HPV type, or copy number of the tumor and the grade of differentiation or keratinization or the clinical stage. After a median follow-up of 13 months, five of nine HPV-positive patients were alive without recurrence, whereas all seven HPV-negative patients had died because of disease. The results of this study indicate a possible major role of HPV in the development of vaginal cancer.

Expression of epidermal growth factor receptor and HER-2/neu in normal and neoplastic cervix, vulva, and vagina.

Monoclonal antibodies were used to localize immunohistochemically epidermal growth factor receptor and HER-2/neu in normal and neoplastic frozen tissue samples from the lower genital tract of women. In squamous epithelia of the cervix, vulva, and vagina, epidermal growth factor receptor and HER-2/neu both were expressed most strongly by basal keratinocytes. Expression of both of these cell surface molecules decreased as cells underwent differentiation toward the mucosal surface. In contrast, both epidermal growth factor receptor and HER-2/neu were expressed throughout the entire thickness of the epithelium by undifferentiated squamous cells in squamous metaplasia, raised condyloma, and carcinoma in situ. In 34 squamous cancers of the cervix, vulva, and vagina, all malignant cells were found to have moderate to heavy staining for epidermal growth factor receptor. Staining of 33 of these cancers for HER-2/neu was light, although one patient who presented with distant metastases had heavy staining for HER-2/neu. These data suggest that although overexpression of HER-2/neu in squamous cancers of the lower genital tract is a rare event, it may be associated with aggressive biologic behavior.

Tumor markers in gynecologic neoplasms.

The applications of tumor markers and steroid receptors in gynecologic neoplasms are described. The value of immunohistologic techniques in the histogenetic assessment of gynecologic neoplasms is examined. Approaches to the diagnosis of similar-appearing lesions are presented.

Immunohistochemical and electron microscopic findings in benign fibroepithelial vaginal polyps.

Eleven classic benign "fibroepithelial polyps" of the vagina were examined using a panel of immunocytochemical agents. Two were also examined electron microscopically. In all cases the stellate and multinucleate stromal cells characteristic of these lesions stained strongly for desmin, indicating muscle intermediate filament production. In common with uterine fibroleiomyomata, numerous mast cells were also often seen. Myoglobin staining was negative. Electron microscopical examination confirmed that the stromal cells contained abundant thin filaments with focal densities and also showed the ultrastructural features usually associated with myofibroblasts. It is concluded that these tumours would be better designated polypoid myofibroblastomas in view of the above findings.

Vulvovaginal polyps. Histologic appearance, ultrastructure, immunocytochemical characteristics, and clinicopathologic correlations.

A clinicopathologic study of 18 vulvovaginal fibroepithelial polyps with a comparison to normal stroma is presented. Twelve cases were analyzed by immunohistochemical methods for the presence of vimentin, desmin, muscle-specific actin, myoglobin, S-100 protein, alpha 1-antitrypsin (AAT), alpha 1-antichymotrypsin (ACHT), cytokeratin (AE1/AE3), and epithelial membrane antigen. Stromal cells reacted with vimentin antiserum in eight cases. Desmin reactivity was detected in five of 12 cases, four of which coexpressed vimentin. Two cases exhibited muscle-specific actin reactivity, and a single case weak AAT reactivity. The stromal cells were unreactive with S-100 protein, epithelial membrane antigen, cytokeratin, ACHT, and myoglobin. Ultrastructurally, the stromal cells of four fibroepithelial polyps resembled fibroblasts and myofibroblasts. Our immunohistochemical and ultrastructural data suggest that the stromal cells of fibroepithelial polyps are a collection of functional fibroblasts that may be capable of differentiating along two or more cell lines. Dramatically elongated cytoplasmic processes extend from both normal vulvovaginal stromal cells and the cells of the polyps. The cell attachments to each other and to bundles of collagen suggest a potential for a physiologic role in tissue contractility, especially in the immediate postpartum state. The common association with pregnancy may represent a local exuberant response to some presently unidentified trophic or stimulating factor.

[Postoperative pseudosarcomas of the genitourinary tract. A diagnostic trap. Presentation of 4 cases of which 2 were studied immunohistochemically and review of the literature]. / Pseudosarcomes post-opératoires du tractus génito-urinaire. Un piège diagnostique. Présentation de 4 observations dont 2 avec étude immunohistochimique et revue de la littérature.

Four cases of postoperative pseudosarcomatous nodules of the genitourinary tract are reported. Two occurred in the vagina of 38 and 57 year-old women, 1 and 3 months after vaginal hysterectomy. The third one occurred in the bladder of a 60-year-old man, 1 and 1/2 months after prostatic adenomectomy. The fourth one occurred in the bladder of a 60-year-old woman, 2 months after transurethral cystoscopy with biopsies. On microscopic examination, the lesions were cellular spindle cell proliferations with numerous mitoses and minimal or no cytonuclear atypia. They mimicked sarcoma, with an overall appearance suggestive of leiomyosarcoma. Immunohistochemical studies performed on 2 cases showed strong reactivity of the cells with vimentin. Cytokeratins, desmin, smooth muscle myosin, smooth and striated muscle actins and factor VIII were negative. Despite simple, sometimes partial, local excision of the nodules, there was neither recurrence nor metastasis 8 months to 5 years later.

Correlation of histology with human papillomavirus DNA detection in the female genital tract.

Genital condyloma and intraepithelial neoplasia secondary to Human Papillomavirus (HPV) infection are characterized by perinuclear halos and marked nuclear atypia (koilocytotic atypia) on cytologic and histologic examination. However, at times the histologic findings, including the degree of nuclear atypia, may be suggestive but not absolutely diagnostic of an HPV related neoplasm. HPV DNA sequences were detected in 63 and 56% of colposcopically visible vaginal and cervical lesions, respectively, that were diagnosed as condyloma or intraepithelial neoplasia. HPV DNA sequences were detected in 14 and 47% of vaginal and cervical lesions, respectively, that did not fulfill the histologic criteria of condyloma or intraepithelial neoplasia (i.e., "nondiagnostic"). When examining cervices from patients with no visible lesion and no recent history of an abnormal pap smear, 5.5% had detectable HPV DNA sequences. The histologic findings in this group were equivalent to the virus-negative cases and similar to the "nondiagnostic" cervical lesions. These findings suggest that the detection rate of HPV DNA in "nondiagnostic" tissues is dependent on the site and presence or absence of a visible lesion. The rate is similar in cervical lesions regardless of the histologic findings whereas it is less in vaginal lesions when the histologic criteria of condyloma or intraepithelial neoplasia are not detected.

DNA analysis cervical and vaginal cancer cells during radiotherapy by rapid high-resolution cytometry.

The DNA content of the nuclei of cancer cells of 12 cases of cervical cancer and 2 cases of vaginal cancer, treated with radiotherapy, were studied in 50 specimens. Specimens were taken from each case before radiotherapy and at the totals of 1,000 rad, 2,000 rad, 3,000 rad and 4,500 rad (or 5,000 rad). All specimens were stained by the Papanicolaou method and were analyzed by rapid high-resolution cytometry. Total optical density, mean nuclear area and the 5N-exceeding rate (5NER) increased gradually following irradiation. Cancer cells disappeared in good response cases before 3,000 rad. Eight smears with a 5NER under 100 at the dose of 3,000 rad or more seemed to be poor response cases. Low 5NER and low mean nuclear areas were observed in both patients who died with persistent disease after radiotherapy, as well as in one case treated with chemotherapy for persistent disease after radiotherapy.

Squamous cell carcinoma antigen for monitoring cervical cancer.

The tumour-associated antigen was determined in the plasma of patients with squamous cell carcinoma (SCC) of the uterine cervix by radioimmunoassay. Setting a limit of 2 ng/ml, levels were abnormal in 13.4% of healthy controls, in 14% of patients with carcinoma in situ and in 62% of patients with invasive cervical SCC. The incidence of elevated SCC antigen levels and the absolute antigen plasma concentration were dependent upon the tumour load, increasing significantly with advanced stage disease. Abnormal SCC antigen values in operable cervical cancer declined to normal within one week after radical hysterectomy with pelvic lymphadenectomy. In cases of radiotherapy antigen values took 4-6 weeks after the start of treatment to return to normal. The success of both treatment modalities was announced by an early rise in the SCC antigen in the initial phase of therapy, followed by normalisation. After successful primary treatment and a complete remission during further follow-up SCC antigen in plasma was only increased in 3.8% of the cases. Retrospective evaluations in ten patients with progressive disease showed the reappearance of abnormal SCC titers and further increase preceeding the clinically detectable relapse or progression, with a median interval of 8 weeks. The present study indicates that SCC antigen determination is not useful for the early diagnosis of cervical cancer, but it is a potential means for monitoring the efficacy of individual anticancer therapy of SCC of the uterine cervix and for detecting recurrent disease.

Comparison of fresh and paraffin-embedded tissue as starting material for DNA flow cytometry and evaluation of intratumor heterogeneity.

Flow cytometric analysis of DNA ploidy and S-phase fraction (SPF) from paraffin-embedded tumors has become an important diagnostic and prognostic tool in clinical pathology and investigative oncology. The present study aimed at elucidating the reliability of the method. About 90% of the 1,400 paraffin-embedded tumors analyzed were evaluable for DNA index and about 70% for SPF, although considerable differences between various tumor types were detected. The within-assay coefficients of variation for determination of tumor DNA index and SPF were 2% and 6%, respectively. Intratumor variation in DNA index was observed in 24% of breast and in 21% of ovarian carcinomas and variation in SPF in 36% and 29% of the respective tumors. Intratumor variation in SPF was greatest in DNA-diploid tumors, which may indicate that SPF values in these tumors may be less reliable owing to variations in the proportions of tumor and nontumor cells. If the methodological variation and the intratumor variation were taken into account, there was a good correlation between DNA indices (r = 0.980) and between SPF values (r = 0.794) obtained from fresh and paraffin-embedded tumors. It is concluded that accurate determination of DNA index and SPF from paraffin-embedded tumors is possible in the majority of cases. Regardless of the type of starting material used for DNA flow cytometry it is advantageous to study several samples from each tumor to account for the intratumor heterogeneity.

Nuclear DNA content of clear cell adenocarcinoma of the vagina and cervix and its relationship to prognosis.

A study was performed to determine if ploidy levels of nuclear DNA content could be used as a prognostic index of the biologic behavior of clear cell adenocarcinoma of the vagina and cervix in young females. Forty tumors were examined. Four patterns of nuclear DNA distribution were identified: (1) peridiploid stem cell lines (2N to 3N); (2) peritetraploid stem cell lines (3N to 5N); (3) hypertetraploid stem cell lines (greater than 5N); and (4) highly aneuploid without detectable stem cell lines. Tumors having peridiploid and peritetraploid stem cell lines (low ploidy group) were most often clinical Stage I or II (87%) and were in general associated with a favorable prognosis. Tumors having hypertetraploid stem cell lines and highly aneuploid distribution (high ploidy group) were usually clinical Stage III or IV (65%). Clinically advanced tumors (Stage III or IV) had poor prognosis irrespective of ploidy level. Among the clinical Stage I and II tumors, the low ploidy group had a slightly better prognosis than the high ploidy group. This difference, however, was not statistically significant.

Identification of human papillomavirus DNA in cervical and vaginal intraepithelial neoplasia with molecularly cloned virus-specific DNA probes.

The presence of human papillomavirus (HPV) DNA was identified in the tissues of cervical and vaginal intraepithelial neoplasia by Southern blot DNA hybridization under conditions of low stringency. The specific types of HPV present in the tissues were identified by using molecularly cloned types 1 through 6 (HPV-1 through HPV-6) HPV DNA probes under high-stringency conditions. All tissues of cervical and vaginal intraepithelial neoplasia analyzed contained HPV genomes. Fifteen of 19 samples (79%) contained HPV-6 DNA, and 10 of 19 samples (53%) HPV-3 DNA. Hybridization with HPV-1, HPV-2, HPV-4, and HPV-5 DNAs was also observed in several of the samples. Four of the samples did not hybridize with any of the probes tested (HPV-1 through HPV-6); yet, all showed hybridization with an HPV-EV DNA (a type 3-related DNA) probe under low-stringency conditions, indicating the presence of HPV types other than those belonging to HPV-1 through HPV-6.

Keratin polypeptide composition as a biochemical tool for the discrimination of benign and malignant epithelial lesions in man.

An investigation was undertaken of the keratin polypeptides of various benign and malignant tumors of the human skin and vaginal epithelium by one and two dimensional gel electrophoresis. The keratin patterns of benign tumors were found to be similar to the patterns of normal epithelium or stratum corneum. The relative proportion of stratum-corneum associated keratin polypeptides to those polypeptides characteristic for the living layers corresponds to morphological features (e.g., hyperkeratosis, acanthosis). In contrast to benign tumors, epithelial carcinomas totally lack the group of high-molecular-weight keratins. This finding may be helpful in the diagnostic discrimination between benign and malignant epithelial lesions.