宇田川 康博

We review the cases of 31 patients with stage IVb or recurrent cervical adenocarcinoma who were treated with combination chemotherapy utilizing mitomycin C, etoposide, and cisplatin (MEP). The total response rate was 16.1% (95% confidence intervals (CIs), 5.5 to 33.7%) with 4 patients having a complete response (CR) and 1 having a partial response. In patients with no prior chemotherapy, the response rate was 26.7% (95% CIs, 7.8 to 55.1%) with 2 of these CR patients surviving over 3 years, 1 a disease-free survival, A marked response was found in distant recurrent lesions. The major toxicity was myelosuppression. Forty-five percent of patients I-rad leukocytopenia above grade 3; thrombocytopenia and anemia were not common. In patients with cervical adenocarcinoma and no prior chemotherapy, there was a moderate response to MEP therapy. (C) 1999 Academic Press.

Objective. Ovarian clear cell carcinoma is commonly associated with pelvic endometriosis. We retrospectively evaluated clinicopathological data on the association between ovarian clear cell carcinoma and pelvic endometriosis. Methods. Between 1984 and 1995, we evaluated clinicopathological data on 53 Japanese patients with primary ovarian clear cell carcinoma who had been initially treated at Keio University Hospital. The clinical backgrounds and 5-year survival rates were evaluated. Results. Twenty (37.7%) of the 53 patients had carcinoma accompanied by pelvic endometriosis. These 20 cases were classified as FIGO stage I (n = 13, 65%), stage II(n = 1, 5%), stage III(n = 6, 30%), or stage IV (n = 0). The other 33 cases of ovarian clear cell carcinoma had no evidence of association with endometriosis and were classified as stage I (n = 19, 57.6%), stage II (n = 2, 6.1%), stage III (n = 9, 27.2%), or stage TV (n = 3, 9.1%). The incidence of a positive intraperitoneal cytology in stage Ic was significantly less in the group with endometriosis than in that without the endometriosis (n = 1, 14.3% vs n = 9, 64.3%, P = 0.03). The 5-year survival rate of stage I patients was significantly greater in ovarian clear cell carcinoma with pelvic endometriosis (100%) than in that without it (60%, P < 0.05). Conclusion. Patients having ovarian clear cell carcinoma with pelvic endometriosis exhibited a better prognosis than those without endometriosis, especially those patients with stage I cancer. (C) 1999 Academic Press.

Employing RMG-1 cells derived from human ovarian adenocarcinoma and COS-1 cells transfected with cDNA for beta 1,4-galactosyltransferase, we first investigated the intracellular localization of this enzyme anal secondly assayed separately the two types of soluble forms of it (designated as normal GalT and GAT) in their culture supernatants. As results, the binding of anti-beta 1,4-galactosyltransferase monoclonal antibody was strongly positive at the Golgi area surrounding nuclei and the staining pattern was the same between RMG-1 cells and COS-1 transformants. These data demonstrated that the cells were producing beta 1,4-galactosyltransferase and had the ability to release it into the culture medium. As for the two soluble farms released from the cells, they were simultaneously detected in the culture medium by Western blotting and enzyme immunoassay using monoclonal antibodies, i.e., Mab8628 reactive to both GAT and normal-GalT and Mab8513 specific for GAT, indicating that both were derived from RMG-1 and from the cDNA introduced into COS-1 cells. Since serum-GAT clinically reflects tumor growth more accurately or specifically than normal GalT does, RMG-1 and COS-1 cells having cDNA for beta 1,4-galactosyltransferase would be promising as sources of these enzymes in order to investigate the differences in the behaviors of normal GalT and GAT associated with ovarian malignant tumors.

SKG-3a and SKG-3b are two distinct human uterine cervical epidermoid carcinoma cell lines derived from a single donor. We studied these two closely related cell lines from the standpoint of drug susceptibility. The growth inhibitory effects of cisplatin (CDDP), doxorubicin (ADM), etoposide (VP-16), and paclitaxel (taxol) on SKG-3a and SKG-3b cells assessed by crystalviolet dye uptake assay were almost the same. SKG-3b cells treated with CDDP, ADM, VP-16, and taxol showed the apoptotic cell death, whereas apoptosis in SKG-3a cells was not induced by these anticancer drugs. Caspase-3 activity was increased only in the SKG-3b cell lysate after treatment with CDDP, ADM, and VP-16 but was not found in the SKG-3a cell lysate. These results indicate that despite growth inhibitory effects of anticancer drugs being almost the same, there may be differences in the common signaling pathways involved in the apoptotic process between SKG-3a and SKG-3b obtained from the same tumor.

Clinically relevant animal models of human cancer are important for studies of cancer biology, invasion and metastasis, acid for investigating new forms of prognostic diagnosis and therapy. An ovarian tumor line (RMG-1: human clear cell carcinoma of the ovary) previously grown subcutaneously was implanted orthotopically as intact tissue into the ovarian capsule of 22 nude mice. The tumors showed progressive growth at the orthotopic site in all animals. Tumor-associated serum galactosyltransferase (GAT) tended to be positive in all nude -mice. The tumors invaded or metastasized to the contralateral ovary, retroperitoneum, mesentery and peritoneum, and omentum, and metastasized to the subcutaneous tissue, lymph nodes and distant organs including the liver, kidney, pancreas, and diaphragm, In striking contrast, subcutaneous transplantation of this tumor resulted in growth in only 2 of 5 animals,vith local lymph node and kidney involvement but no retroperitoneal or peritoneal involvement. These findings suggest that orthotopic implantation provides a suitable micro-environment in which ovarian cancer can express its intrinsic clinically-relevant properties. This approach is relevant to the clinical features of ovarian cancer and is thought to be a useful model for studies of therapy for this cancer.

In this study clinical studies were conducted on galactosyltransferase associated with tumour (GAT) as a newly developed marker of ovarian cancers. The positive rates of GAT with a cut-off value of 16 U/ml (which corresponds to the mean + 2 standard deviations (S.D.) for healthy females) were 4.7% for benign ovarian tumours, 4.5% for endometriosis and 45.9% for ovarian cancers. GAT showed a positive rate comparable to that of CA546 or CA72-4 among other tumour markers (CA602, CA125, CA546, CA72-4, STN and SLX) examined in ovarian cancers. However, it showed lower positive rates for benign ovarian diseases and, in particular, it gave the lowest positive rate for endometriosis among the aforementioned tumour markers. Furthermore, the receiver operating characteristic (ROC) analysis for discriminating between ovarian cancer and endometriosis showed a significantly high area under the curve (AUC) for GAT compared with that of the other markers. GAT showed the lowest correlation coefficients with other markers, and the positive rate and the diagnostic efficiency were increased by its combination assay with CA602 and/or CA546. Furthermore, the accuracy of the diagnosis of ovarian cancer improved by examining GAT after screening with CA602 or ultrasonography. These results suggest that GAT is a suitable marker for distinguishing ovarian cancers from benign gynaecological diseases, particularly endometriosis, and is useful for combination assay or secondary screening for ovarian cancers. (C) 1998 Elsevier Science Ltd. All rights reserved.

Objective: Dienogest, a synthetic steroid with progestational activity, is used as a component of oral contraceptives and is currently being evaluated clinically for the treatment of endometriosis. Thee present study was conducted to confirm the effects of dienogest on experimental endometriosis in rats and to elucidate its mechanism of action. Design: Experimental endometriosis induced by autotransplantation of endometrium in rats. Methods: Endometrial implants, immune system, and bone mineral were investigated after 3 weeks of medication. Results: Dienogest (0.1-1mg/kg per day, p.o.) reduced the endometrial implant volume to the same extent as danazol (100 mg/kg per day, p.o.). Simultaneously, dienogest ameliorated the endometrial implant-induced alterations of the immune system: i.e. it increased the natural killer activity of peritoneal fluid cells and splenic cells, decreased the number of peritoneal fluid cells, and decreased interleukin-1 beta production by peritoneal macrophages. In contrast, danazol (100 mg/kg per day, p.o.) and buserelin (30 mu g/kg per day, s.c.) had none of these immunologic effects. Additionally, combined administration of dienogest (0.1 mg/kg per day) plus buserelin (0.3 mu g/kg per day) suppressed the bone mineral loss induced by buserelin alone, with no reduction of the effect on endometrial implants. In vitro studies on dienogest revealed an antiproliferative effect on rat endometrial cells due to inhibition of protein kinase C activity plus a partial progestational effect. Conclusions: Dienogest appears to be a potent agent with mechanisms of action different from those of danazol and GnRH agonists currently available for the treatment of endometriosis.

A monoclonal antibody (MSN-3) was raised using HEC-108 cells derived from poorly differentiated endometrial carcinoma as the immunogen. The immunoglobulin subclass of MSN-3 was IgGr1. The target antigen of MSN-3 was a protein with a molecular weight of 77 kDa, and it was shown to be localized in the cytoplasm. MSN-3 only reacted with 14% of normal proliferative endometrium cells, but it showed a high positivity rate of 66% for endometrial carcinoma. The target antigen of MSN-3 increased as endometrial cells became more malignant, and the possibility of changes in localization was also suggested. Moderately and poorly differentiated endometrial carcinoma showed a high positivity rate for MSN-3. MSN-3 reacted rarely or not at all with normal cervical glandular tissue, but the positivity rate for cervical adenocarcinoma (especially endocervical adenocarcinoma) was a high rate of 59%. The patterns of staining of endocervical adenocarcinoma by MSN-3 included diffuse staining of the whole cytoplasm and not only that near the glandular lumen, as well as staining of the basal cytoplasm. Changes in the localization of the target antigen were clearly associated with carcinogenesis of the cervical glandular cells. The MSN-3-positive rate was high in patients with lymph node metastasis and vascular invasion. Among the staining patterns, the basal and diffuse patterns tended to increase with malignacy. The basal pattern of staining was characteristic of MSN-3, suggesting that it might assist in the diagnosis of cervical adenocarcinoma.

We examined the expression of beta 1,4-GT gene products in 11 gynecological cancer cell lines. A 4.7 kb mRNA and protein (54,000 Da and 57,000 Da) were detected by Northern blot and Western blot. Immunocytochemical staining revealed that beta 1,4-GT was localized in the Golgi or ER of tumor cells. An intense beta 1,4-GT mRNA signal was detected in ovarian and cervical cancer cells, whereas the level of beta 1,4-GT mRNA was very low in uterine endometrial cancer cells. We also confirmed that expression of beta 1,4-GT mRNA corresponded to expression of beta 1,4-GT protein. These results suggest that expression of the beta 1,4-GT gene products is higher in human cervical and ovarian cancer cells than in uterine endometrial cancer cells.

Changes in CA602 and CA546, two carbohydrate-related antigens newly established in our institute, were evaluated during the course of in-vitro fertilization and embryo transfer, The concentration of CA602, an antigen with characteristics similar to those of CA125, did not change substantially until embryo transfer, In patients with clinical evidence of ovarian hyperstimulation syndrome (OHSS), a significant increase in serum concentrations of CA602 was observed 10 days after embryo transfer (P < 0.01), There were no significant changes in the patients without OHSS. A significant elevation of oestradiol concentration in OHSS patients was also observed, with the variation before oocyte retrieval being greater than that seen with CA602, The concentration of CA546, which had lower false-positive rates in benign disease, did not differ significantly in the OHSS and non-OHSS patients, Our data demonstrate that serum concentrations of CA602 are increased even in early stages of OHSS, and suggest a possible role for this antigen as a marker for OHSS.

Dienogest is an orally active synthetic steroid that is used for contraception and is currently being studied for the possible treatment of endometriosis. Earlier we demonstrated that dienogest had therapeutic effects on experimental endometriosis in rats and that its mechanisms of action were different from those of drugs currently on the market for the treatment of endometriosis. We also reported preclinically that dienogest showed a potential anticancer action against hormone-dependent cancers that was different from that of progestins. Accordingly we obtained preclinical background data for the above-described clinical applications and extension of the clinical use of the drug in the near future by investigating the endocrinological profile of dienogest in rabbits and rats. Dienogest was characterized by having a moderate binding affinity for progesterone receptors and by progestational activities: it stimulated endometrial proliferation (greater than or equal to 0.01 mg/kg) that was only partially inhibited by RU-486, and induced carbonic anhydrase activity in endometrium (greater than or equal to 0.01 mg/kg). Also, it was slightly uterotrophic (greater than or equal to 1 mg/kg) with very low binding affinity for oestrogen receptors and slightly antioestrogenic (greater than or equal to 1 mg/kg). In addition, dienogest showed slight binding to androgen receptors but without biological androgenic and anabolic activities (100 mg/kg), with neither glucocorticoid activity nor mineralocorticoid activity (100 mg/kg), and with very slight binding affinity for human sex hormone-binding globulin. These findings suggest that dienogest is not a pure progestin and appears to induce fewer side effects than drugs currently on the market for the treatment of endometriosis.

We have developed a new procedure for the selective determination of β1-3 and β1-4 galactosyltransferases with Lc3Cer as the substrate and the microsomes of fetal and adult porcine livers as the enzyme sources. This method was based on the detection of such products as Lc4Cer for β1-3 galactosyltransferase (β1-3GT) and nLc4Cer for β1-4 galactosyltransferase (β1-4GT), with monoclonal anti-Lc4Cer and anti-nLc4Cer antibodies, respectively. This method thus enabled us to differentiate the activity of β1-BGT from that of β1-4GT with a high degree of sensitivity. The method was then used to determine the activities of both enzymes in human gynecological carcinoma-derived cells. Four of the five cell lines derived from uterine endometrial cancer expressed significantly high levels of specific activity of β1-3GT among the cell lines examined, while their β1-4GT activities were less than 20% of that for β1-3GT in the endometrial carcinoma-derived cells. On the other hand, a higher specific activity of β1-4GT than that of β1-3GT was detected in the cell lines derived from uterine cervical and ovarian cancers. These findings were thus found to correlate closely with the rate of expression of Lc4Cer- and nLc4Cer-based carbohydrate chains in the cell lines based on the results staining.