Combination assay of urinary beta-core fragment of human chorionic gonadotropin with serum tumor markers in gynecologic cancers.

Ectopic production of the immunoreactive beta-subunit of human chorionic gonadotropin (IR-hCG beta) by gynecologic malignancies has been well recognized, but IR-hCG beta has not yet been established as a clinically useful tumor marker, except for germ cell tumors. We measured the concentrations of IR-hCG beta-related molecules, intact hCG, free hCG beta, and beta-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-hCG beta was obtained in the assay for beta-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 beta-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 beta-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 beta-CF. All of the ovarian cancer patients examined were detected by the combination assay of urinary beta-CF and serum CA125. The levels of urinary beta-CF showed little correlation with those of the serum tumor markers, indicating the usefulness of the combination assay of urinary beta-CF with serum tumor markers for detecting cervical and ovarian cancers.

Purification and characterization of protein kinase C from a higher plant, Brassica campestris L.

Protein kinase C (PKC) was partially purified from Brassica campestris L., by successive chromatographies on DEAE-cellulose membrane, hydroxyapatite and phenyl-5PW columns. The purified preparation showed typical characteristics of the conventional type of mammalian PKC that responds to Ca2+, phosphatidylserine, and diacylglycerol or the tumor-promoting phorbol ester, phorbol 12-myristate 13-acetate. The plant PKC activity was apparently associated with a 75-kDa polypeptide that was recognized by an antibody against the catalytic domain of rat PKC. Substrate specificity of the plant PKC was similar to that of the rat PKC. A synthetic peptide corresponding to residues 4-14 of myelin basic protein, which is a selective substrate for the mammalian PKC, was phosphorylated efficiently by the plant PKC. These results indicate the existence of a PKC equivalent in higher plant cells.

[Assessment of urinary beta-core fragment of hCG as a tumor marker of cervical cancer].

Beta-core fragment (beta-CF), a fragment of the hCG beta-subunit missing its carboxyterminal peptide, can be detected in the urine of women throughout pregnancy or in trophoblastic disease. It is also found in the urine of patients with nontrophoblastic cancers. We examined the beta-CF level in urine samples from patients with cervical cancer and assessed its value as a tumor marker. beta-CF was measured by an enzyme immunoassay with hCG beta-core directed monoclonal antibody No. 229. Based on the cut-off value (0.2ng/ml) from control subjects, the overall positivity rate for urinary beta-CF in the cervical cancer group was 45% (57 of 128 patients), increasing from 32% (23 of 73) in stage I to 100% (2 of 2) in stage IV. These positivity rates exceeded or equaled those of the other markers, SCC, CEA, CA19-9 and CA125, simultaneously measured in the patients' serum. There was no significant difference between the positivity rates for the two histological types of cancer, squamous cell carcinoma and adenocarcinoma. Serial determination in 28 patients with increased urinary beta-CF prior to therapy showed that 24 patients had a decreased concentration after successful treatment, but 2 of 4 patients with still increased urinary beta-CF during or after treatment subsequently relapsed. The determination of urinary beta-CF may provide a useful tool in monitoring the response to treatment in patients with cervical cancer.

[Investigation of 5-fluorouracil (5-FU) levels and pyrimidine nucleoside phosphorylase activities in the tissues from patients with uterine cervical and ovarian cancers after oral administration of 5'-DFUR].

The usefulness of 5'-DFUR in both patients with uterine cervical and ovarian cancers was investigated by determining pyrimidine nucleoside phosphorylase (PyNPase) activities and 5-FU levels in cancerous and normal tissues resected from them after oral administration of 5'-DFUR. In uterine cervical cancer, each group of 9 cases administered single dose of 400 mg of 5'-DFUR and 7 cases administered 400 mg of 5'-DFUR 3 times a day continuously for 7 days was investigated. In ovarian cancer, all of 9 cases were administered 400 mg of 5'-DFUR 3 times a day continuously for 7 days. In conclusion, PyNPase activities in the tissues of uterine cervical and ovarian cancers were higher than those in the normal tissues. 5-FU tissue levels in the cancerous tissues were significantly higher than in the normal tissues and blood as well. This tendency was observed in each of the single and continuous administration groups. These results suggest that the tumor selectivity which is one of characteristics of 5'-DFUR could be expected also for cancer in the field of gynecology.

[Distribution of lymphocyte subsets in regional lymph nodes in uterine cervical cancer and its immunological significance].

We performed an immunohistochemical study on the distribution of lymphocyte subsets in the regional lymph nodes in uterine cervical cancer. The lymph nodes from patients with cervical cancer were stained with monoclonal antibodies to Leu 1,2,3 and 7 as the indicators of pan T, killer/suppressor T, helper/inducer T, and NK/K cells, respectively. On the lymph nodes without tumor involvement, there were a greater number of lymphocytes of each subset in the nodes in stage Ib without nodal metastasis than in the nodes in stage Ia and stage Ib with nodal metastasis and stage II, suggesting that the regional lymph nodes have active immune reactions in stage Ib cancer without nodal metastasis. On the lymph nodes with tumor involvement, there were great numbers of lymphocytes of each subset, especially Leu 7+ cells, in the nodes with local invasion, suggesting that very strong immune reactions occur in these nodes. On the other hand, there were very few lymphocytes in the nodes with diffused tumor. The present findings suggest that regional lymph nodes in cervical cancer play important roles in antitumor immune response and, furthermore, even the tumor-involved lymph nodes are of great immunological significance if the metastatic tumor is localized.

[Characterization of immunoreactive hCG beta-subunit in cultured fluids of the cell lines derived from gynecologic malignant tumors].

We characterize the hCG beta-like materials in cultured fluids from 12 different cell lines derived from gynecologic malignant tumors (SKG-1, -2, -3a and -3b cervical squamous carcinoma, SNG-M and -2 endometrial adenocarcinoma, SKN uterine sarcoma, RTSG ovarian undifferentiated adenocarcinoma, RMUG ovarian mucinous adenocarcinoma, RKN ovarian sarcoma, RMG ovarian clear cell carcinoma and NJG gestational choriocarcinoma) by three kinds of enzyme immunoassay (EIA) which were specific for whole hCG, free hCG beta and beta-core fragment, respectively. Of eleven nontrophoblastic cell lines, nine secreted hCG beta-like immunoreactive substance. The above-mentioned three EIAs for each fractionated specimens with gel chromatography on Sephadex G-100 revealed that the immunoreactivity in the SKG-2 and RTSG cultured fluids were totally attributable to free hCG beta but neither to whole hCG nor beta-core fragment. On the other hand, the NJG choriocarcinoma cell line secreted both whole hCG and free hCG beta, no beta-core fragment could be detected in the cultured fluid. The present results suggested that ectopically produced hCG beta-like material may represent the free hCG beta molecule, and that the beta-core fragment may not be a cellular secretory product.

[Electron microscopic and immunological studies concerning the effect on the antitumor activity of sizofiran (SPG) combined with radiotherapy for cervical cancer].

It has been well known that Sizofiran (SPG) cultured from the Schizophyllum commune Fries activates the macrophages and induces the cytotoxic lymphocytes in some cancers. In this study, we observed electronmicroscopically the macrophages around the cancer tissue from the patients with uterine cervical cancer after the treatment with SPG. At the same time, their immune responses were also examined by analyzing lymphocyte subsets, ADCC and NK activity in peripheral blood. A considerable number of erratic macrophage with well developed Golgi apparatus, endoplasmic reticulum and mitochondria were found in the uterine cervical cancer tissue from the patients treated with SPG under radiotherapy. Simultaneously, we identified the lysosome granules with a bright filament structure which appeared to be specific for SPG. In the immune responses evaluated by analyses of peripheral blood, the number of CD 16+ cells and NK activity significantly increased in the patients treated with SPG as compared with non-treated group. The present results indicate that SPG-immunotherapy combined with radiotherapy not only induces the cytotoxic activity of macrophage but also augments NK activity in the patients with uterine cervical cancer.

[Clinicopathological study on microscopic urinalysis as screening for malignant cells].

Study on detection of malignant cells in urinary sediments using supravital staining was described. We examined 96,554 specimens of urinary sediments for 2 years from January 1985 to December 1986. The results of microscopic urinalysis were compared with the cytological and histological diagnoses. Atypical cells were found in 138 patients, and 47 (34.1%) cancers were diagnosed histologically among them. These included 33 bladder cancer, 1 ureter cancer, 1 renal pelvic cancer, 2 prostate cancers, 1 rectal cancer, and 9 uterine cancers. Seven patients of them had not been under suspicion of malignancy yet before atypical cells were detected. Therefore microscopic urinalysis caused the triggers of cancer diagnoses. For bladder cancers, the positive rates in microscopic urinalysis were 43.4%, and those in urinary cytology were 52.4%. The positivity revealed higher in high-grade cancers than in low-grade. As compared with the results between microscopic urinalysis and urinary cytology in identical patients, the rate of correspondence between them was 89.5%. In 61.2% of positive and suspicious urinary cytology, atypical cells were not found. Atypical cells were seen in negative urinary cytology of 26 cases, and 5 cases of cancers were diagnosed histologically. These suggested that microscopic urinalysis as a screening for malignant cells was useful to detect urinary tract malignancy combining with urinary cytology.

[Cytological and histological effects of regional hyperthermia and radiation in cancer of the uterine cervix].

Hyperthermia is known to produce tumor-specific effect. Its combination with radiotherapy has been found to enhance antitumor effect and hyperthermia is a new modality of cancer treatment. Here, we report a cytological and histopathological study of the effects of hyperthermia on uterine cervical cancer. Our subject cases were 63 patients with cervical cancer (squamous cell carcinoma). Thirteen cases were treated with hyperthermia. Of these, the clinical stage was I b in 2, stage II in 10, stage III in 1. Before radical operation, external irradiation was given at 40Gy. with Lineac. During this period, the lesion was treated with hyperthermia at 42-45 degrees C for 30-60 min. with a 13.56 MHz RF capacitive heating system "Endoradiotherm 100A" (Kureha Chemical Industry CO. Ltd.) and an intraluminal applicator designed for uterine cervix. The control cases received radiotherapy alone before radical operation. There were 15 cases in Stage I b, 19 in stage II, and 16 in stage III. After the completion of radiotherapy, radical hysterectomy was performed in all cases. The effects of irradiation on the lesion were compared cytologically and histologically for different doses. The evaluation of the irradiation damage in cancer cells in the cytological study was based on findings such as intracytoplasmic vacuolation, the formation of giant nuclei, intranuclear vacuolation and formation of polynuclei. The degree of irradiation damage was classified into 5 grades from Do (little irradiation effect) to Dx (no viable cancer cells remaining), and the irradiation effect in the histopathological study was classified in to 8 grades from Grade 0 to Grade IVC according the Oboshi and Shimosato's classification. Cytological changes at 10Gy. were rated as D3 (relatively extensive irradiation damage in cancer cells) in 38.5% of the hyperthermia group, as compared to about 20% in the control groups. The cases rated Grade II B histologically made up 38.5% of the hyperthermia group: this grade was about 10% in the control groups. This suggests that radiation was more effective in the hyperthermia group than in the control groups, as was also suggested by the cytological results. With irradiation at 30Gy., cytological changes were rated as Dx in 61.5% of the hyperthermia group, as compared to 25.0-26.7% of the control groups. Histological results were Grade III, with only cancer cells regarded as non-viable observed, in 38.5% of the hyperthermia group and Grade IV, with no cancer cells, in another 23.1%. In the control groups, there were no cases with Grade IV, and those with Grade III totalled 12.5-26.7%.(ABSTRACT TRUNCATED AT 400 WORDS)

[Immunologic significance for regional lymph node histology in uterine cervical cancer].

To evaluate the immunological significance for regional lymph node histology in uterine cervical cancer, 1,144 lymph nodes of 153 cases were studied. Four histological findings in lymph nodes as follows were evaluated: Paracortical Area (PA), Sinus Histiocytosis (SH), Germinal Center (GC) and Degenerative Changes (DC). PA was well developed in early cancer, while hypoplastic in advanced cancer. SH appeared in early stages and disappeared gradually as cancer progressed. GC was most hyperplastic in stage I b and hypoplastic in stage I a and III. DC was remarkable in advanced cancer. The findings of PA of the regional lymph nodes in the cases without metastasis, and those of PA, GC and DC of the metastatic lymph nodes in the cases with metastasis were well correlated with prognosis. These results indicate that investigation of the histology of lymph nodes draining uterine cervical cancer may give valuable information to evaluate the immune response of the patients and to predict their prognosis.

[Cytologic studies of cervical adenocarcinoma].

Seventy three cases of cervical adenocarcinoma, classified histologically according to WHO classification, were cytodiagnostic examined for cellular arrangement, cellular shape, nuclear shape, chromatin pattern and appearance rate of nucleolus. In adenosquamous carcinoma, the observed cancer cells were classified further into five categories and the incidence of each type was clarified. The authors obtained the following conclusions. The cytological findings in cellular arrangement, cellular shape, chromatin pattern and nucleolus findings were closely related with the degree of differentiation of endocervical type adenocarcinoma. Since no substantial cytodiagnostic differences were seen between endometrioid adenocarcinoma and endocervical type adenocarcinoma, it appears to be difficult to differentiate between them. Clear cell adenocarcinoma is characterized by sheet-like cell arrangement, polycrystal cell shape, round nuclei, finely granular chromatin pattern and appearance of macronucleolus. Among them the occurrence of macronucleolus was clearly observed and appeared of high value in making a diagnosis. Adenosquamous carcinoma, showing no characteristic finding in cytology, is proved to have morphological diversity. Careful observation of the intermediate type cancer cells is necessary to presume the histology of adenosquamous carcinoma.