Kazuo Hasegawa

Expression of α and β genes of human chorionic gonadotropin in lung cancer

To confirm the ectopic production of human chorionic gonadotropin (hCG) in lung cancer, we attempted to detect the presence of mRNA transcripts of the α and β genes for hCG in lung cancer tissues obtained from surgical operations. Although we were able to show the presence of hCGβ mRNA transcripts in lung cancer tissue by Northern blot, the sensitivity of the assay was too low for a precise analysis of hCGB mRNA transcripts in most lung cancers. Using reverse transcription PCR (RT‐PCR) and Southern blot analysis, however, various amounts of mRNA transcripts of hCGβ genes 3, 5, 7 and 8 were demonstrated in 9 of the 14 lung cancer tissues examined, while no mRNA transcripts were detectable in 12 normal lung tissues from the same patients. Our results are consistent with a clear difference in serum and urinary hCGβ levels observed between normal subjects and lung cancer patients. The expression of the hCGα gene, however, was detected in normal lung tissues more frequently than in lung cancer tissues using RT‐PCR Southern blot. Our results strongly suggests the production of hCGβ as being part of the phenotype of malignantly transformed lung cells and further strengthen its superior specificity over intact hCG or hCGα as a tumor marker for lung cancers. Int. J. Cancer 71:539‐544, 1997.

Irinotecan (CPT-11) and Cisplatin as First-Line Chemotherapy for Advanced Ovarian Cancer

Objective: To evaluate the efficacy and toxicity of a combination of irinotecan (CPT-11) and cisplatin as first-line chemotherapy in advanced ovarian cancer. Methods: Twenty-six patients with previously untreated advanced epithelial ovarian cancer were enrolled in this study. CPT-11 60 mg/m2 was administered intravenously on days 1, 8, and 15 in combination with cisplatin 60 mg/m2 on day 1. Cycles were repeated every 28 days for at least two cycles. The median patient age was 55 years (range, 37–75), and the median performance status was 1. Results: Objective responses were recorded in 19 of 25 eligible patients (76%; 95% confidence interval, 55–91%). Complete responses were obtained in 2 patients (8%), and partial response in 17 patients (68%). Stable disease was recorded in 2 patients (8%) and progressive disease in 2 (8%). The median time to response was 62 days (range, 28–234 days). The median survival time for all 25 patients was 30.9+ months (range, 4.1–60.0+ months). The major toxic effects were leukopenia, neutropenia, and diarrhea. Grade 3 or 4 leukopenia, neutropenia, and diarrhea occurred in 17 (68%), 20 (83.3%), and 5 patients (20%), respectively. Thrombocytopenia was less common. No treatment-related deaths occurred. Conclusion: The combination of CPT-11 and cisplatin showed significant activity in chemotherapy-naive patients with advanced ovarian cancer. Neutropenia was the dose-limiting adverse effect, whereas diarrhea was mainly mild to moderate.

Mutational Analysis of BRCA1 Gene in Ovarian and Breast-ovarian Cancer Families in Japan

We analyzed the alteration of BRCAI in DNA obtained from 83 individuals of 13 Japanese site‐specific ovarian cancer families and 6 breast‐ovarian cancer families. Six germline mutations were detected in 7 families, which consisted of 4 breast‐ovarian cancer and 3 site‐specific ovarian cancer families, by single‐strand conformation polymorphism analysis, followed by direct sequence determination. The mutations included three framcshifts, two nonsense mutations, and one missense mutation causing loss of a zinc‐binding motif. The frequency of loss of heterozygosity at the microsatellite markers on the BRCAI gene was 57% (8 of 14 cases) in site‐specific ovarian cancer families, and 100% (6 of 6 cases) in breast‐ovarian cancer families. All tumors of the patients carrying a mutation of BRCAI showed deletion of wild‐type alleles, implicating BRCAI as a tumor suppressor gene. Tbese results suggest tbat germline mutations of the BRCAI gene play an important role in the carcinogen‐esis of breast and/or ovarian cancer in a majority of breast‐ovarian cancer families and in some site‐specific ovarian cancer families.

Immunotherapy using the streptococcal preparation OK-432 for the treatment of uterine cervical cancer. Cervical Cancer Immunotherapy Study Group.

The effectiveness of immunotherapy using a streptococcal preparation, OK-432, was evaluated for cervical cancer. The 382 eligible patients were stratified by presence/absence of surgical operation and clinical stage, and then, in each stratum, were randomly divided into two groups: an OK-432 treatment group and a control treatment group. The 3-year recurrence-free rates of 221 patients in the OK-432 group and 161 patients in the control group were 71.9% and 58.6%, respectively. The intergroup difference was statistically significant (P less than 0.05). Delayed skin reactions to phytohemagglutinin (PHA) and Su-polysaccharide extracted from Streptococcus pyogenes Su-strain (Su-PS) and peripheral lymphocyte counts were reduced within two months after the initiation of therapy in both groups. The observed immunological changes were apparently reversed by 3 months after the start of the therapy in the OK-432 group, but this took at least one year in the control group, with significant intergroup differences at 6 and 12 months of the therapy (P less than 0.01). These results indicate that OK-432 can be considered as one of the most effective and useful immunotherapeutic agents for cervical cancer.

Molecular Forms of Human Chorionic Gonadotropin in Choriocarcinon Serum and Urine

The molecular forms of human chorionic gonadotropin (hCG) were assessed in parallel serum and urine samples from a choriocarcinoma patient by gel chromatography, isoelectric focusing, and Western blot analysis. There were significant qualitative differences in the molecular forms of hCG between the serum and the urine. The serum hCG had a greater molecular weight and a stronger acidic charge than the urinary hCG. These results may be attributed to differences of sialic acid contents. Affinity chromatography using Ricinus communis agglutinin‐conjugated Sepharose showed that the serum hCG was completely sialylated, while the urinary hCG was partially desialylated. In addition to the complete hCG molecule, two kinds of hCGβ‐related fragments were detected in the urine of the patient, but not in the serum. In the Western blot using anti‐hCGβ or anti‐hCGβ‐CTP under reducing conditions, the urine presented a new band (CTP′) with Mr 23,000 in addition to the β‐subunit, while the serum revealed a single band of the β‐subunit with or without a reducing reagent. The present data revealed striking qualitative differences in the molecular forms of hCG between serum and urine of a patient with choriocarcinoma.

Screening of BRCA1 mutation using immunohistochemical staining with C-terminal and N-terminal antibodies in familial ovarian cancers

We examined the subcellular localization of BRCA1 proteins using immunohistochemical staining with C‐terminal (GLK‐2 antibody) and N‐terminal (Ab‐2 antibody) monoclonal antibodies in 44 familial ovarian cancers. Among these, 24 cases were associated with 13 independent germ‐line mutations of BRCA1, and loss of heterozygosity (LOH) at one or more BRCA1 microsatellite markers was found in all 21 informative tumors tested. With GLK‐2 antibody, cytoplasmic staining was observed in 15 of 16 tumors (93.8%) with mutation in exon 11, and BRCA1 staining was absent in 8 of 8 tumors (100%) with mutation in exons other than exon 11. When immunohistochemical staining was performed with Ab‐2 antibody, both nuclear and cytoplasmic staining were observed in 14 of 16 tumors (87.5%) with mutation in exon 11. Interestingly, nuclear staining was observed in 3 of 3 tumors (100%) with mutation downstream of exon 11, even though no staining was detected in 5 of 5 tumors (100%) with mutation upstream of exon 11. On the other hand, in familial ovarian cancers without BRCA1 mutations, nuclear staining or both nuclear and cytoplasmic staining was observed in 18 of 20 specimens (90%) and 20 of 20 specimens (100%) with GLK‐2 antibody and with Ab‐2 antibody, respectively. These results suggest that an immunohistochemical assay in combination with employing the C‐terminal and the N‐terminal antibodies appears to have potential as a reliable and useful technique for the screening of BRCA1 mutations, at least to predict the status of mutation, upstream or downstream of exon 11.

Ovarian thecoma with ascites and high serum levels of CA125

Abstract We report a 34-year-old woman with an ovarian thecoma and ascites who exhibited high serum levels of CA125. Measuring serum tumor markers and imaging are two important diagnostic tools for malignant ovarian tumors. In the present case, a preoperative diagnosis of benign ovarian tumor could not be made due to the elevation of CA125 (895 U/ml) and nonspecific MRI findings.

Combination Assay of Urinary β-Core Fragment of Human Chorionic Gonadotropin with Serum Tumor Markers in Gynecologic Cancers

Ectopic production of the immunoreactive β‐subunit of human chorionic gonadotropin (IR‐hCGβ) by gynecologic malignancies has been well recognized, but IR‐hCGβ has not yet been established as a clinically useful tumor marker, except for germ cell tumors. We measured the concentrations of IR‐hCGβ‐related molecules, intact hCG, free hCGβ, and β‐CF, in the sera and urine of patients with various gynecologic cancers (cervical, endometrial, and ovarian cancers) to assess their clinical usefulness as a tumor marker in comparison with serum tumor markers such as CEA, SCC, CA125, and CA19‐9. The highest incidence of IR‐hCGβ was obtained in tbe assay for β‐CF in the urine, with positive rates of 47.7% (94 of 197) for cervical, 37.8% (14 of 37) for endometrial, and 84.4% (38 of 45) for ovarian cancers with a cut‐off value of 0.2 ng/mg of creatinine. In cervical cancer, there was no significant correlation between the concentrations of urinary β‐CF and serum SCC, and 57.9% (114 of 197) of the patients were detected by the combination assay of these tumor markers. Serial determination in 22 cervical cancer patients with elevated urinary β‐CF level prior to therapy showed that its level decreased after successful treatment, but 4 of 5 patients with persistent or recurrent disease had elevated levels of urinary β‐CF. All of the ovarian cancer patients examined were detected by the combination assay of urinary β‐CF and serum CA125. The levels of urinary β‐CF showed little correlation with those of the serum tumor markers, indicating the usefulness of the combination assay of urinary β‐CF with serum tumor markers for detecting cervical and ovarian cancers.

Use of Localization and Activity of Thymidine Phosphorylase in Human Gynecological Tumors for Predicting Sensitivity to Pyrimidine Antimetabolite Therapy: An Observational Study

Background: Thymidine phosphorylase (dThdPase) is the rate-limiting enzyme in the conversion of 5 ′ -deoxy- 5-fluorouridine (5 ′ -dFUrd), an intermediate metabolite of capecitabine (Xeloda ® ), to 5-fluorouracil (5-FU). We investigated the correlation between dThdPase activity and immunohistochemical staining in gynecological carcinoma and adjacent normal tissues. We hypothesize that the differential dThdPase activity between tumors and adjacent tissue is predictive of response to treatment with pyrimidine antimetabolites. Methods: In 45 samples of carcinoma tissue and 35 of adjacent normal tissue from 45 patients, we measured dThdPase activity as well as immunoreactivity using an anti-dThdPase monoclonal antibody and macrophage and histiocyte-specific antibodies. Results: dThdPase activity in tumor tissue was significantly higher than that in the corresponding adjacent normal tissue in all samples tested (12 uterine cervical, 19 endometrial, and 4 ovarian tumors). Anti-dThdPase immunopositivity was observed in the epithelial tumor cells of 76.9% of uterine cervical cancer samples, 60.0% of endometrial cancer samples and 63.6% of ovarian cancer samples. In stromal tissue, 84.6% of uterine cervical tumors (11/13), 90.0% of endometrial tumors (18/20), and 81.8% of ovarian tumors (9/11) were immunopositive for anti-dThdPase in interstitial cells (mainly macrophages). Macrophages were also strongly reactive in the stromal tissues of uterine cervical, endometrial, and ovarian cancers. The correlation between dThdPase activity and intensity of immunohistochemical staining of epithelial tumor cells with anti-dThdPase monoclonal antibody was statistically significant in endometrial carcinoma ( P = 0.008) but borderline in uterine cervical tumors ( P = 0.077). We found a good correlation between dThdPase activity and staining of epithelial tumor cells, particularly in the case of endometrial cancer. Conclusions: We show that gynecological carcinomas show increased dThdPase activity, and this activity correlates with dThdPase staining of tumor epithelial cells. Thus, dThdPase staining of biopsy specimens could be useful in predicting the outcome of therapy with pyrimidine metabolites.

Paclitaxel-cisplatin combination in advanced ovarian cancer: a phase II study

AbstractBackground. To date there have been four large randomized studies in Western countries examining the role of combining cisplatin and paclitaxel as first-line treatment for ovarian cancer. A phase II study of paclitaxel and cisplatin in Japanese patients with advanced ovarian cancer was performed to determine the objective response rate and toxicity of this regimen. Methods. Previously untreated patients with stage III or IV ovarian cancer and a good performance status were eligible. Treatment consisted of paclitaxel 180 mg/m2 administered as a 3-h intravenous (i.v.) infusion followed by cisplatin 60 mg/m2 i.v. Treatment was repeated every 3 weeks for at least four cycles. Granulocyte colony-stimulating factor (G-CSF) was not routinely used. Results. Among 26 eligible patients, there were 4 complete and 17 partial responses, for an overall response rate of 80.8% (95% confidence interval [CI], 60.6% to 93.4%). One hundred and twenty-nine treatment cycles were administered to the 26 patients. Grade 4 neutropenia was observed in 64 treatment cycles (50%) and in 23 patients (88%). Thrombocytopenia was less common. The most common nonhematologic toxicities included neurotoxicity, fatigue, arthralgia/myalgia, and nausea/emesis. Conclusion. Paclitaxel plus cisplatin is a highly active regimen in patients with advanced ovarian cancer. The toxicities of this regimen are well tolerated.