Temporal associations of human papillomavirus infection with cervical cytologic abnormalities.

The relationship between infection with different human papillomavirus types and cervical intraepithelial neoplasia was studied in a group of 398 women seen in a private gynecology practice in Washington, D.C. Each woman was assessed for human papillomavirus infection by Southern blot hybridization analysis of cervical cells obtained by swab. The human papillomavirus results were correlated with the results of Papanicolaou smears taken the same day and with data abstracted from medical records regarding past cervical disease. Subjects with normal cytologic findings at the time of human papillomavirus testing were followed up for an average of 2 to 3 years with additional Papanicolaou smears. At the time of human papillomavirus testing, 58% (19/33) of women with cervical intraepithelial neoplasia had detectable human papillomavirus deoxyribonucleic acid in contrast to 10% (28/289) of women with normal cytologic findings (p less than 0.001). This association persisted after statistical adjustment for age and current use of oral contraceptives, a factor that appeared to increase the detection of human papillomavirus. Among women with no current cytologic evidence of neoplasia, human papillomavirus detection was more likely in those with a history of past genital neoplasia (p = 0.05). In the follow-up study, 15% (3 of 20) of cytologically normal women who were human papillomavirus-positive at baseline subsequently exhibited cervical cells suggestive of cervical intraepithelial neoplasia compared with only 5% (9 of 195) of human papillomavirus-negative women. However, this difference reflected recurrent and not incident neoplasia.

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.

Detection of pS2 messenger RNA in gynecological cancers.

Estrogen-inducible pS2 mRNA was previously detected in human cancer cell lines the growth of which was sensitive to estrogen. In the present study, the expression of the pS2 gene was analyzed in 111 gynecological carcinomas. The pS2 message was detected in greatest abundance in 6 primary carcinomas of the ovary (6 of 29), 4 of these being mucinous cystadenocarcinomas. A secondary carcinoma of the ovary, and another of the omentum (1 of 4), also contained detectable levels of pS2 mRNA. Weak pS2 mRNA signals were occasionally observed in endometrial (2 of 55) and cervical carcinomas (2 of 33) as well. There was a poor correlation between estrogen receptor and pS2 mRNA in ovarian carcinomas.

Procoagulant activity of mononuclear phagocytes from different anatomical sites in patients with gynecological malignancies.

Mononuclear phagocytes exhibit complex interactions with cancer cells and might contribute to fibrin formation associated with malignancy through the production of procoagulant activity (PCA). We have studied the PCA of peritoneal macrophages in 8 patients with advanced (stages III or IV) ovarian cancer and of macrophages from regional lymph nodes in 14 patients with limited (stages I or II) uterine cancer; peritoneal and lymph-node macrophages from patients with benign gynecological tumors were used as reference cell populations. In all patients, PCA of blood monocytes was also studied. Peritoneal and lymph-node macrophages obtained from patients with ovarian and uterine cancer, respectively, expressed far higher levels of basal PCA than the corresponding cell populations from patients with benign tumors (p less than 0.001). PCA of blood mononuclear cells from patients with ovarian, but not with uterine cancer, was significantly higher (p less than 0.001) than that of control cells. High levels of D-dimer, a specific product derived from plasmin-induced degradation of stabilized fibrin, were found in all ascitic fluids and in all plasma samples but one from patients with ovarian cancer. In contrast, all controls and all uterine cancer patients but one had normal plasma D-dimer. Our findings suggest that local activation of host macrophages for PCA production might contribute to fibrin formation within the tumoral mass. In advanced cancer, blood monocytes may also be activated to produce PCA and thus contribute to activation of intravascular coagulation and, possibly, to thrombo-embolic complications frequently associated with disseminated malignancy.

[The cellular localization of the fertility alpha 2-microglobulin in tumors of the reproductive system]. / Kletochnaia lokalizatsiia al'fa-2-mikroglobulina fertil'nosti v opukholiakh reproduktivnoi sistemy.

Seventy-five samples of human tumors were examined immunohistochemically for fertility alpha-2-microglobulin also known as progesterone-associated protein of the endometrium. The protein was detected in 35.7% (5 of 14) endometrial cancer samples and in 20% (2 of 10) of ovarian malignancies. Tumors of the stomach, colon, breast, lung and some other sites failed to reveal the antigen with the exception of a single case of pulmonary adenocarcinoma which showed the ectopic expression of the protein. Fertility alpha-2-microglobulin is stage-specific tissue marker of the endometrium. It is expressed at the final stage of cell differentiation and is partially lost in cancer.

[Extramammary Paget's disease. Immunocytochemical study and histogenetic considerations]. / Malattia di Paget extramammaria. Studio immunocitochimico e considerazioni istogenetiche.

We report a case of extramammary Paget's disease arising in the anogenital region in association with an underlying sweat gland microcarcinoma. Paget's cells were investigated with monoclonal anti-EMA and anti-cytokeratin antigen and with mono- and polyclonal anti-CEA antigen. Positive immunostaining was observed in Paget's cells and in the underlining tumor, whereas keratinocytes and melanocytes did not stain. CEA was also detected in cells and secretions of normal apocrine glands. The immunohistochemical use of polyclonal anti-keratin and anti-S-100 protein antigen is helpful in differentiating Paget's disease from other morphologically similar skin lesions, such as Bowen's disease and superficial spreading melanoma in situ.

An immunohistochemical study of neuroendocrine cells in gynecologic tumors.

Various gynecologic tumors with argyrophilia were studied immunohistochemically for chromogranin using two antibodies, antichromogranin and antineuroendocrine. Of seven small cell carcinomas of the cervix, four were immunoreactive with antichromogranin and seven with antineuroendocrine. Argyrophil cells of six cervical adenocarcinomas were all immunoractive with both antibodies. Type I argyrophil cells of 20 endometrial carcinomas were likewise stained positively. However, of the 30 endometrial carcinomas with type II argyrophil cells, 19 showed positive immunoreactivity for chromogranin and 22 for neuroendocrine. Of the ovarian tumors tested, argyrophil cells of 11 mucinous tumors, three carcinoid tumors, and the pancreatic tissue of a malignant mixed germ cell tumor were all chromogranin- and neuroendocrine-immunoreactive. Type I argyrophil cells of five endometrioid carcinomas of the ovary were also immunoreactive with both antibodies. Of the 13 endometrioid carcinomas with type II argyrophil cells, only four showed positive immunoreactivity for chromogranin and only five for neuroendocrine. In conclusion, both antichromogranin and antineuroendocrine detect the specific neuroendocrine markers in close association with argyrophilia in gynecologic tumors, the latter being more sensitive for small cell carcinoma of the cervix, and for type II argyrophil cells in adenocarcinoma of the endometrium and endometrioid carcinoma of the ovary.

[Correlation of carcinoembryonic antigen (CEA) and serum and tissue alpha-fetoprotein (AFP) in malignant tumors of the female genital tract]. / Correlazioni tra antigene carcinoembrionario (CEA) ed alfa-fetoproteina (AFP) sierico e tissutale nei tumori maligni della sfera genitale femminile.

The Authors have carried out determinations of AFP in the serum and of CEA in serum and in surgically removed tissue of 67 patients with genital tract's malignant tumors. As the results obtained among values of pre and post operatory CEA and tissue concentrations are incongruous, they believe useful, within the monitoring of patients with gynecological cancer, dosing CEA in the serum and also on the surgical section trying to reduce so the minimum positive false and negative false. Finally the repeated dosage of these antigens allows to follow the local replaced or the metastasis after some month, before the symptoms are clear.

CA 125 in normal tissues and carcinomas of the uterine cervix, endometrium and fallopian tube. I. Immunohistochemical detection.

The distribution of cancer antigen 125 (CA 125) has been investigated in normal tissues and carcinomas of the Müllerian duct by immunohistochemical methods using the monoclonal antibody OC 125. Detection of CA 125 was most intense in cryostat sections and decreased in formalin fixed and paraffin embedded tissues according to the duration of fixation. Enzymatic digestion with neuraminidase or alkaline hydrolysis abolished specific staining suggesting the antigen is a sialylsaccharide bound to protein by alkali-labile linkage. Immunohistochemical staining demonstrated the presence of CA 125 in all normal glandular epithelia of the endocervix, endometrium and fallopian tube in different distribution patterns. In normal endometrium the cellular distribution pattern was related to the menstrual cycle. In endocervical, endometrial and tubal adenocarcinomas CA 125 was found in 73% of cases. In glandular structures the antigen was concentrated at the luminal surface of the tumour cells, in solid tumour areas it was spread throughout the cytoplasm or concentrated in large cytoplasmic vacuoles. The expression of CA 125 was considerably lower in solid tumour areas. These data show that CA 125 is not a true "tumour marker", but a product of female genital mucosae and of their cancerous derivates provided their synthesizing ability is not lost in the course of pathologic differentiation.

CA 125 in normal tissues and carcinomas of the uterine cervix, endometrium and Fallopian tube. II. Immunoradiometric determination in secretions, tissue extracts and serum.

The study deals with the occurrence of cancer antigen 125 (CA 125) in the normal and neoplastic uterine cervix, endometrium and fallopian tube and its applicability as a tumour marker. CA 125 concentrations were measured in 52 secretion specimens, in cytosol fractions of 97 tissue biopsies and in serum from 47 women with nonmalignant disorders and from 334 patients with carcinomas. High quantities of CA 125 (780-454860 U/ml) were detected in cervical mucus, intra-uterine and tubal fluid, exceeding those in the corresponding serum samples by factors of up to 2000. CA 125 concentrations were 9-53 fold higher in cytosol fractions of normal and neoplastic glandular epithelia of the endocervix and endometrium than in those of cervical squamous epithelia and the cervical wall. Despite similarly high antigen concentrations in normal glandular epithelia and adenocarcinomas serum levels elevated to above 65 U/ml were only found in patients with malignant tumours. The positivity rates in serum increased with tumour extent and were 0-43% for primary and 63-79% for recurrent cervical, endometrial and tubal adenocarcinomas. During long-term follow-up, CA 125 serum concentrations were concordant with the clinical course in 10 out of 11 patients with progressive carcinomas. According to these results, the release of CA 125 into the peripheral blood is apparently dependent on the infiltrative growth and the mass of the tumour rather than on the local tissue concentrations. The clinical use of CA 125 is limited to the detection of advanced adenocarcinomas of the Müllerian duct.

Epidermal growth factor receptor expression in gynecological malignancies.

Epidermal growth factor receptor (IEGF-R) levels were analyzed in 72 gynecological tumor specimens. Measurable EGR-R levels were found in a significant percentage of ovarian and uterine tumors. Moreover, all vulvar epidermoid carcinomas and uterine sarcomas analyzed were EGF-R positive. In all tumor types examined, scattered EGF-R levels were observed. Higher EGF-R levels were found in metastatic than in primary ovarian tumors. Moreover, EGF-R were found to be more expressed in less differentiated than in well-moderately differentiated endometrial tumors. Our results suggest a role of EGF or EGF-like substances in regulating the growth of gynecological malignancies, and indicate EGF-R expression as a possible prognostic factor.

Comparison of formalin, buffered formalin, and Bouin's fixation on the detection of human papillomavirus deoxyribonucleic acid from genital lesions.

Detection of nucleic acid sequences homologous to human papillomavirus (HPV) relies primarily on their extraction from unfixed tissue. We detected HPV sequences in DNA extracted from paraffin-embedded tissue fixed in formalin (buffered and unbuffered) and Bouin's solution by dot blot hybridization. A detectable hybridization signal was noted in 32% of these fixed tissues which were chosen from cases where HPV DNA was detected in the unfixed tissue. When using a homologous 32P-labeled probe and a high stringency wash, the hybridization signal was lost if DNA was extracted after Bouin's fixation and diminished after formalin fixation, more so with unbuffered formalin. Similar differences in the hybridization signals among the different fixatives after high stringency wash were noted with in situ hybridization. Southern blot analysis showed that DNA extracted from tissues fixed in Bouin's was degraded and ranged in size from 100 to 500 base pairs as compared with 100 to 900 base pairs for DNA extracted from tissue fixed with unbuffered formalin. In contrast, no degradation was noted after fixation with buffered formalin. These results demonstrate that HPV sequences can be identified in DNA extracted from paraffin-embedded, fixed tissue. However, use of some fixatives may preclude identification of HPV type, by either dot blot or in situ hybridization.

Detection of human papillomavirus DNA in anogenital exophytic lesions by in situ hybridization to paraffin sections.

In situ hybridization was used to detect human papillomavirus (HPV) nucleic acids (type 6b, 11, 16, 18, 31, and 33) and immunohistochemistry was done to detect papillomavirus common antigen in paraffin sections of biopsy specimens. Two patients suffering from condyloma acuminatum contained HPV-6b and HPV-11. Both cases showed small foci of the antigen-positive cells. One patient having condyloma acuminatum with dysplastic features contained a small quantity of HPV-16 without any antigen-positive cells. One case of verrucous carcinoma showed neither HPV-DNA nor antigen. In situ hybridization is a powerful tool in the analysis of the pathogenesis of HPV-associated neoplasms.

Analysis of tumor heterogeneity in a patient with synchronously occurring female genital tract malignancies by DNA flow cytometry, DNA fingerprinting, and immunohistochemistry.

A case of a patient with bilateral ovarian cancer and a uterine malignant mesodermal mixed tumor with ascites and metastatic disease is presented. Flow cytometry, DNA fingerprinting, and immunohistochemistry were performed to assess the origin of these malignancies. Ploidy analysis showed that both ovarian tumors had different aneuploid stemlines (DNA index [DI] = 1.64, 1.85, right ovary and DI = 1.73, left ovary) indicating independent origins. One of the stemlines in the right ovary (DI = 1.64) was also present in the ascites cells, whereas omentum metastases showed the same stemline (DI = 1.73) as the left ovarian tumor. The uterine malignancy contained three aneuploid stemlines. The highest stemline was associated with epithelial differentiation, but a metastatic origin from the left ovarian tumor seems unlikely. DNA fingerprinting analysis revealed a common change in restriction fragment length pattern in the DNA from all tumor localizations as compared with the patient's constitutional DNA. These results indicate that DNA flow cytometry can be helpful in discriminating intragenital metastatic disease from multiple primary tumors.

Revision de linfaticos de la pelvis y su relacion con el cancer ginecologico / Pelvic female lymphatics and their relationship with gynecologic cancer

Se revisaron las cadenas linfaticas de la pelvis femenina y el drenaje linfatico de los organos sexuales femeninos, haciendose enfasis en analizar las vias de extension del cancer en los diferentes organos, y la patofisiologia de la extension del Cancer pelvico femenino

[The place of tumor markers in breast and gynecologic neoplasms]. / Place des marqueurs tumoraux dans les néoplasies mammaires et gynécologiques.

The main tumor markers used in breast and gynaecological neoplasias are reviewed with their individual characteristics: carcino-embryonic antigen (CEA), alpha-fetoprotein (AFP), chorionic gonadotropin hormone (CGH), CA 125, CA 15.3 and Squamous Cell Carcinoma Antigen (SCC). Their indications are specified according to the following cancer locations: breast, trophoblast, ovary, endometrium and cervix. The major clinical applications of the tumor markers routinely used are emphasized.

Antigenic markers of genital carcinoma.

No significant testing in screening or early diagnosis of cancer is possible at present. Tumor markers of different origin are useful in the search for occult locoregional recurrence, metastatic disease, or early detection of progression irrespective of localization. The comparative analysis of tumor markers for follow-up is mandatory. Estrogen- and progesterone-receptor assays for prognostic evaluation in endometrial and breast carcinoma, and in some cases of ovarian tumors, should complete any pretherapeutic step. There is a possible link with epidermal-growth factor (EGF) with regard to uncontrolled growth of tumors in hormone-dependent (target) organs.