Protooncogene amplification and tumor ploidy in human ovarian neoplasms.

DNA from 24 ovarian tumors, including 16 carcinomas, was examined for amplification of the proto-oncogenes c-myc, int-2, and rc-erbB-2. All cases of carcinoma were also examined by flow cytometry for DNA ploidy and cell cycle analysis, and eight cases of carcinoma were examined for estrogen and progesterone receptors. Protooncogene amplification was not detected in the DNA of benign ovarian neoplasms, or of ovarian carcinomas with low malignant potential. Amplification of c-myc was detected in six of 12 cases of invasive carcinoma, int-2 amplification was present in one case, and c-erbB-2 amplification was not detected in any case. Among the seven cases evidencing protooncogene amplification, three cases showed aneuploidy in tumor DNA, while four showed diploidy. Two cases which showed aneuploidy in tumor DNA did not demonstrate any degree of protooncogene amplification. Protooncogene amplification was frequently associated with morphologic nuclear anaplasia and high mitotic count. Six of the seven cases demonstrating c-myc or int-2 were of the serous type or showed some degree of serous differentiation, while none of the four cases of purely mucinous carcinoma had evidence of amplification. While the total number of cases in the study was limited, it would appear from the trend demonstrated by the data that protooncogene amplification (particularly c-myc) may be involved in the pathogenesis of aggressive common epithelial tumors of the ovary.

Serous papillary adenocarcinoma of the endometrium. Analysis of proto-oncogene amplification, flow cytometry, estrogen and progesterone receptors, and immunohistochemistry.

Primary and metastatic tumor tissues of serous papillary adenocarcinoma of the endometrium were examined for the following: (1) amplification of int-2, c-erbB-2 and c-myc proto-oncogenes by Southern blot hybridization; (2) DNA ploidy by flow cytometric study; (3) and expression of specific proteins, such as estrogen and progesterone receptors, keratin, vimentin, and carcinoembryonic antigen (CEA) using immunohistochemical and biochemical techniques. Amplification of c-myc was observed in the specimens from the endometrium (ten-fold) and from omental metastasis (five-fold). Both int-2 and c-erbB-2 amplification were not observed. The tumor showed aneuploidy, with the specimens from the endometrium and omental metastasis exhibiting multiple populations of aneuploid tumor cells. Estrogen and progesterone receptors could not be detected biochemically; however, immunohistochemically, estrogen receptors were observed in tumor cells forming papillary structures but not in the tumor cells of the solid, more poorly differentiated areas. A similar distribution was observed for both low and high molecular weight keratin. The findings of c-myc amplification and aneuploidy in the serous papillary adenocarcinoma of the endometrium are consistent with its aggressive behavior observed clinically and emphasize the importance of distinguishing this lesion from other types of endometrial carcinoma.

Tumor markers of epithelial ovarian neoplasms.

Ninety-three formalin-fixed, paraffin-embedded surgical specimens from 69 ovarian tumors representing all five epithelial cell types were studied by immunohistochemistry, peanut and ulex lectin binding, and carbohydrate histochemistry. Carcinoembryonic antigen (CEA) was mostly noticeable in mucinous tumors (21 of 26). Glycogen was highly prevalent in clear cell (8 of 9) and endometrioid (4 of 6) carcinomas, in contrast to serous carcinomas (3 of 6), where it was only focally distributed, and was completely absent in all mucinous tumors. Among the different types of malignant tumors examined, mucinous carcinomas most frequently contained neutral mucins (6 of 8). In mucinous tumors, an increase in CEA content and a decrease in the total mucin secretion, particularly the strongly acidic sulfated group, were found to parallel the increased malignant potential of the tumor. Peanut and/or ulex lectin binding was a feature common to almost all epithelial neoplasms. Although peanut lectin showed a slightly higher affinity to serous and clear cell tumors, while ulex lectin was bound more to mucinous and endometrioid neoplasms, distribution of D-galactose and L-fucose does not have a diagnostic utility in these tumors. Placental lactogen was detected in 3 of 17 benign tumors and one of 19 tumors of low malignant potential (LMP). The beta subunit of hCG was found in one of 17 benign tumors, in 2 of 19 LMP tumors, and in 3 of 31 carcinomas.

[A study of localizations and serum data; four tumor markers (CA125, CA19-9, CEA and TPA) in ovarian cancers].

Four tumor markers (CA125, CA19-9, CEA, TPA) were analysed between the localizations and the serum data in the 36 ovarian cancers. The positive rates of each tumor markers were; 23 cases (63.9%) on CA125, 15 cases (41.7%) on CA19-9, 12 cases (33.3%) on CEA, and 27 cases (75.0%) on TPA. CA125 and CA19-9 were observed in the luminar borders and cellular membranes on serous adenocarcinomas and in the cytoplasms on mucinous adenocarcinomas. Both CEA and TPA were stained in cytoplasms. The correlation in the localization was observed between CA19-9s and CEAs (Kendall's rank correlation = 0.5303 greater than 0.5). The correlations between the localization and the serum data were observed on CA19-9 and CA125.

Immunolocalization of alpha-amylase in ovarian mucinous tumours.

The distribution of alpha-amylase was studied immunohistochemically in 42 cases of ovarian mucinous tumour. Intense immunoreactivity for amylase was found in 6 of 8 cases of mucinous cystadenocarcinoma. In contrast, only 6 of 20 benign mucinous cystadenomas showed immunoreactive amylase, which was weak and patchy. Mucinous cystadenomas of borderline malignancy showed an intermediate degree of amylase immunoreactivity. The patterns seen are very similar to those reported in normal endocervix, cervical glandular atypia, and invasive adenocarcinoma and suggest molecular as well as morphologic similarities in neoplasia at these sites.

[Comparative study on human chorionic gonadotropin receptor in normal human ovary with ovarian tumors].

Human Chorionic Gonadotropin (HCG) is major physiological luteotropic factors for the human corpus luteum. The observations strongly suggest that the human ovary possesses a gonadotropin receptor in the cell membrane. We studied the HCG receptor in normal human ovary and ovarian tumors. Twenty-three human ovarian specimens and 16 ovarian tumor specimens were obtained from women patients having gynecological surgery. Ovaries were homogenized and sonicated. The homogenates were centrifuged at 2000 g for 15 min. After sucrose density gradient ultracentrifugation (78,000 g, 4 h), two fractions were collected from layer of 33% and interface between 33% and 37%. Thirty micrograms of ovarian protein, 8 ng 125I-HCG and unlabeled HCG in a final volume of 0.5 ml of 0.05 mol/L Tris buffer were incubated at 30 degrees C for 2 h. The results were shown in the table.

[Immunohistochemical studies of ras oncogene product p21 in human ovarian tumors].

In the present study, monoclonal antibody rp-28 directed against the ras gene product p21 was used to evaluate ras p21 expression in malignant and benign ovarian tissues. Some ovarian carcinomas (serous cystadenocarcinoma, mucinous cystadenocarcinoma, undifferentiated carcinoma and clear cell carcinoma) demonstrated intense staining of ras p21. In mucinous tumors, both the frequency of ras p21 positive staining and the staining intensity gradually increased with the degree of malignancy. There was no difference in ras p21 expression according to the clinical stages of ovarian carcinomas. In the metastatic lesions, ras p21 staining was rather weaker than in the primary lesions. It is therefore possible that intense staining of ras p21 is associated with the degree of malignancy in some types of ovarian tumors, and that the expression of ras oncogene product p21 is not enhanced with progression and metastasis in such types of ovarian carcinoma. These results suggest that ras oncogene plays an important role in the carcinogenesis of some types of epithelial ovarian tumors.

[Mucin histochemical and immunohistochemical studies of mucinous ovarian tumors].

Mucinous ovarian tumors, (32 benign, 5 borderline, and 30 malignant) were studied by using mucin histochemical staining and immunohistochemical method. Results showed that neutral and acid mucoproteins were demonstrated in these tumors; but their proportion and distribution were different. For instance, sulfuric acid mucoprotein was found in 12/32 (37.5%) of mucinous cystadenomas and in 25/30 (83.3%) of mucinous cystadenocarcinomas (P less than 0.01). Immunohistochemically, colon-ovarian tumor antigen (COTA) was 100% positive in malignant and borderline cases respectively but only 6/32 (18.8%) in the benign. The differences were statistically significant (P less than 0.01). Meanwhile, the differences between COTA staining and HID/AB staining for cystadenocarcinomas were also significant (P less than 0.05). These data suggest that COTA is more sensitive and specific antigen for mucinous ovarian tumors and may be useful for the early detection of malignant changes of mucinous ovarian tumors.

Gastrin-producing ovarian cystadenocarcinoma: sensitivity to secretin and SMS 201-995.

We report a patient with severe peptic ulcer disease and a right ovarian mass that was found to be a gastrin-producing cystadenocarcinoma. Gastrin production by the tumor was stimulated by secretin and inhibited by the long-acting somatostatin analogue SMS 201-995. Following resection of the tumor, serum gastrin levels and the gastrin response to secretin returned to normal. Histologic examination, including Alcian blue staining for mucin and immunoperoxidase staining for gastrin, revealed gastrin at the base and mucin at the apex of the tumor cells. This report demonstrates secretin stimulation and somatostatin inhibition of gastrin secretion from a cell that is apparently not of endocrine origin.

Peritoneal malignant mesothelioma versus serous papillary adenocarcinoma. A histochemical and immunohistochemical comparison.

In order to evaluate adjunctive histologic methods for separating mesothelioma (MM) and serous adenocarcinoma (SC), we studied 28 and 46 respective cases histochemically and immunohistochemically. Ten serous adenocarcinomas arose primarily in extraovarian sites within the abdomen. Diagnoses in each case were established retrospectively by a combination of electron microscopy and clinicopathologic correlation. A panel of antibodies to cytokeratin (CK), epithelial membrane antigen (EMA), B72.3, placental alkaline phosphatase (PLAP), S-100 protein, carcinoembryonic antigen (CEA), Leu M1, CA-125, and amylase (AM) was applied to paraffin sections of each case. Serous carcinoma was reactive for neutral mucins whereas mesothelioma was not; however, only 50% of adenocarcinoma cases stained in this manner. Peritoneal mesothelioma showed reactivity for CK (28 of 28 cases), EMA (24 of 28 cases), AM (five of 28 cases), CA-125 (four of 28 cases), and S-100 protein (three of 28 cases), but lacked B72.3, PLAP, and CEA. Three mesotheliomas expressed Leu M1, but in an extremely focal distribution. Serous carcinoma reacted for CK (46 of 46 cases), EMA (46 of 46 cases), CA-125 (42 of 46 cases), S-100 protein (40 of 46 cases), Leu M1 (34 of 46 cases; with diffuse staining), B72.3 (33 of 46 cases), PLAP (29 of 46 cases), AM (15 of 46 cases), and CEA (six of 46 cases). Two profiles (S-100 + B72.3; S-100 + PLAP) were seen in 41 of 46 serous adenocarcinoma cases but were absent in all mesotheliomas. Hence, these combinations of determinants are effective in separating such neoplasms diagnostically. Moreover, diffuse reactivity for Leu M1, B72.3, PLAP, or CEA in papillary peritoneal neoplasms appears to exclude the possibility of mesothelioma; however, focal Leu M1 reactivity may indeed be seen in mesothelioma. Although CA-125 is a sensitive marker for serous carcinoma, it is not effective in distinguishing it from mesothelioma.

[Quantitative DNA analysis in epithelial ovarian cancer].

The tumor cell nuclear DNA content was measured in 27 cases of epithelial ovarian cancer. The proliferative activity of the tumor was correlated with the prognosis of the patients. The proportion of G0 and G1 cells of the tumors of the same histologic type varied among individuals. There was a close relationship between the proliferative activity of the tumor and the remission rate, duration of survival and mortality of the patients. The heterogeneity of the proliferative activity of the tumor cells may explain the difference of response to chemotherapy. It is suggested that the quantitative analysis of DNA content may be used as an theoretical and experimental basis for the individualization of treatment.

[Distribution of lectin-receptors in ovarian cystadenoma and cystadenocarcinoma].

55 cases of ovarian cystadenoma and cystadenocarcinoma were investigated with a panel of twelve various lectins and ABC technique. Results showed that RCA and WGA reacted with all the tumors, indicating that these two lectins are possibly functional differential markers of both ovarian mucinous and serous tumors. LCA, DBA and SJA might be of considerable help in differential diagnosis of serous cystadenoma and cystadenocarcinoma. PSA probably was a marker indicating malignant change of mucinous cystadenmas. Since there were different reactivities in mucinous and serous cystadenoma, SJA, DBA and SBA might be considered as the functional markers in differentiating these two different types of ovarian cystadenoma.

[Comparative studies on the value of acute phase proteins and CA-125 for monitoring patients with ovarian cancer]. / Vergleichende Untersuchungen über die Wertigkeit von Akute-Phase-Proteinen und CA-125 für das Monitoring von Patientinnen mit Ovarialkarzinom.

In a study of 71 patients with malignant ovarian tumors serum levels of CA-125, C-reactive protein (CRP), alpha-1-antitrypsin and coeruloplasmin were analysed. In contrast to the tumor-free group significantly higher values of CA-125, CRP and alpha-1-antitrypsin were found in the group with recurrent disease. However, the serum-concentrations of coeruloplasmin remained unchanged in both groups. In the group with progressive disease the median values of CA-125 were greater than 65 U/ml and of CRP greater than 12 micron/ml, respectively. The median serum concentrations of alpha-1-antitrypsin (2 to 4 mg/ml) and coeruloplasmin (150 to 600 ng/ml) did not reach their cut-off levels. Beside CA-125 the analysis of CRP and alpha-1-antitrypsin is an additional helpful procedure for the monitoring of patients with malignant ovarian tumors.

Fibrin-fibronectin compounds in human ovarian tumor ascites and their possible relation to the tumor stroma.

Covalently linked heterogeneous fibrin-fibronectin compounds were detected in ascitic fluid of 31 patients with advanced ovarian cystadenocarcinoma by means of enzyme-linked immunosorbent assay techniques, immunoaffinity chromatography, and Western blot analysis. Deposition of fibrin and fibronectin could also be demonstrated immunohistochemically in Carnoy-fixed tissue sections. Fibrin and fibronectin were found in the tumor stroma within tumor nests and more prominently in stroma surrounding the tumor nests. The association of fibrin and fibronectin was especially pronounced in the stroma surrounding the tumor islands. Fibronectin was also found to be associated with stroma cells. Areas within the tumor stroma showed superimposed staining for both fibrin and fibronectin supporting the assumption that the covalently linked fibrin-fibronectin conjugates found in ascitic fluid may stem from the provisional tumor stroma by proteolytic release.

Significance of multiparameter flow cytometric analysis of ovarian cancer.

A prospective flow cytometric examination of ovarian epithelial tumors was undertaken to further characterize aneuploid (including triploid and tetraploid) tumor cell populations according to expression of ovarian tumor antigen CA125 and to expression of class I (normally present in ovarian epithelium) and class II (normally absent in ovarian epithelium) major histocompatibility complex antigens. Samples from thirty-two of 42 patients (76%) exhibited at least one aneuploid population of tumor cells. Separate analysis of the aneuploid and diploid components of samples with aneuploid populations revealed between-tumor variation: seven of 23 aneuploid populations (30%) were positive for CA125; eight of 22 aneuploid populations (40%) exhibited substantial decreases in major histocompatibility complex class I expression, compared with corresponding diploid components of the same samples; and eight of 22 aneuploid populations (36%) exhibited substantial expression of major histocompatibility complex class II antigens. The frequencies of aneuploid populations and of the foregoing antigen expression categories were independent of tumor cell type, stage, and grade. The significance of these results for prognosis remains to be determined.

The potential value of imprint cytology in cytochemical localization of steroid hormone receptors in ovarian cancer.

Steroid hormone receptors were studied in 45 patients with primary, recurrent, or metastatic ovarian cancer in cryostat-frozen sections and imprint preparations. The ligands, 17 B-estradiol-6-carboxymethyloxine-bovine serum albumin fluorescein isothiocyanate (FITC-BSA estradiol) and hydroxyprogesteronehemisuccinate bovine serum albumin tetramethylrhodamine isothiocyanate (TMRITC-BSA progesterone) were used in the fluorescent cytochemical method. Results were compared with standard dextran-coated charcoal (DCC) biochemical assay. An overall significant correlation between biochemical values and cytochemical results was found. However, the imprint results were more sensitive and more specific than the frozen section results. A statistically significant difference (P less than 0.05) was observed between touch preparation material and frozen section specimens by the fluorescent method.

Hormonal activity of epithelial ovarian tumours in post-menopausal women.

Concentrations of progesterone and oestradiol in peripheral serum and tumour cyst fluid were measured in 42 post-menopausal women with epithelial ovarian tumours (17 cancer, 6 borderline malignant, 19 benign tumours) and in 19 post-menopausal women without ovarian neoplasms. The hormonal response of the endometrium was assessed, progestogen and oestrogen receptor content in the tumour tissue case recorded, and tumour deoxyribonucleic acid (DNA) ploidy was measured by flow cytometry. No significant differences were found between the mean serum steroid levels in patients with malignant, borderline or benign tumours, but the mean serum levels of oestradiol in patients with malignant or benign ovarian tumours were higher than those in the controls. Endometrial hormonal activity was seen in 19% of the samples studied. Malignant and benign mucinous epithelial tumours were the types most frequently associated with hormonal activity. Increased levels of sex steroids were seen in the cyst fluid of serous malignant and borderline malignant tumours, while benign tumours were inactive. The steroid receptor content of the various tumour types did not vary significantly. Ten (59%) out of 17 ovarian carcinomas were found to be aneuploid and 41% diploid as measured by flow cytometry. No significant differences in serum levels of progesterone and oestradiol were found between aneuploid and diploid ovarian carcinomas. These results contribute to our knowledge of the hormonal activity of epithelial ovarian tumours in post-menopausal women.

Argyrophilic and hormone immunoreactive cells in normal and hyperplastic pancreatic ducts and exocrine pancreatic carcinoma.

Scattered argyrophil cells were present in normal, large, medium-sized and small pancreatic ducts (ductules). There was marked increase in argyrophil cells in ducts with hyperplastic epithelium. Argyrophil cells were also found in 67.7% of all exocrine pancreatic carcinomas. In a well differentiated group including cystadenocarcinoma, mucinous carcinoma and well differentiated ductal adenocarcinoma argyrophil cells were found in all cases examined. Using four antisera (against insulin, glucagon, somatostatin and gastrin), insulin, glucagon and somatostatin cells were identified in 2.65%, 0.001% and 1.2% of normal ducts, and 7.5%, 2.4% and 4.6% of ducts with hyperplastic epithelium respectively and were also greatly increased in numbers in the latter group. Immunoreactive cells were present in 66.7% of exocrine carcinomas. Cells reactive for insulin were found in 7/15 cases; glucagon in 6/15 cases; somatostatin in 5/15 cases and gastrin in 2/15 cases. Eight cases contained two or more than two types of immunoreactive cells. The presence of argyrophil and hormone immunoreactive cells in pancreatic ducts and carcinomas is indicative of the close developmental relationship between endocrine and exocrine parts of the pancreas. The inter-relationship of response in the different cell types following stimulus suggests that injury to a common precursor may be involved.