Distribution of beta-human chorionic gonadotropin-positive cells in noncancerous gastric mucosa and in malignant gastric tumors.

The authors examined the localization and behavior of beta-human chorionic gonadotropin (HCG)-positive cells in human gastric noncancerous mucosa and in gastric malignant tumors, using immunohistochemistry and the anti-beta-HCG antibody. The beta-HCG-positive cells were located mainly in the antral mucosa and were generally restricted to the neck portion of the pyloric glands, although a few were present in fundic glands of the gastric body. The beta-HCG-immunoreactive cells were found in gastric carcinomas in 53% of the 92 cases examined. These cells were observed more often in advanced carcinomas that were histologically poorly differentiated than in early carcinomas or in well-differentiated tumors, but this prevalence had no statistical significance. The presence of the beta-HCG-positive cells in the gastric carcinomas suggested no appreciable prognostic significance, even quantitatively. In the syncytiotrophoblast-like tumor cells seen in four gastric tumor samples with histologic features of a choriocarcinoma, immunoreactivity to the beta-HCG was striking. There was, however, no recognizable dominance in the number of beta-HCG-reactive cells in the noncancerous mucosa around the tumor.

Establishment and characterization of a continuous in vitro line from a rat choriocarcinoma.

A continuous in vitro cell line of rat choriocarcinoma has been established. It is composed of pure trophoblast cells which multiply and differentiate. The morphology of the cells is very similar to normal rat cytotrophoblasts and giant cells. The cultured cells contain cytokeratin, alkaline phosphatase and express the receptors for Bandeira simplificifolia Agglutinin-I (BSA-I). They are hormonally active as demonstrated by the presence of lactogen and progesterone in the supernatant of the culture. The injected cells develop into choriocarcinoma in syngeneic as well as allogeneic rats. The morphological, biological and immunohistochemical features of these tumors are identical to those described in the transplantable neoplasm from which the in vitro line was established. The presence of Y chromosome in cultured cells proves the paternal origin of the primary tumor developed from extra-embryonic membranes in fetectomized rat and makes this neoplasm similar to human post-gestation choriocarcinoma.

The abnormal occurrence and the differentiation-dependent distribution of N-acetyl and N-glycolyl species of the ganglioside GM2 in human germ cell tumors. A study with specific monoclonal antibodies.

Human primary germ cell tumors were analyzed for the presence of the ganglioside GM2 using three specific monoclonal antibodies which can distinguish the molecular species of the sialic acid moiety: the antibody MK1-16 is specific for N-acetyl GM2, MK2-34 is specific for N-glycolyl GM2, and MK1-17 detects both N-acetyl and N-glycolyl GM2. When the occurrence of the GM2 antigen was tested in 107 cases of human germ cell tumors by the immunohistochemical technique using these antibodies, seminoma was characterized as having the highest frequency of N-acetyl GM2 (89.4%, 42 of 47 cases) among germ cell tumors, followed by embryonal carcinoma (40.0%), and teratocarcinoma (26.6%). Compared with this, yolk sac tumors and choriocarcinoma had a much lower positive incidence of the N-acetyl GM2 antigen. On the other hand, the N-glycolyl GM2 antigen was not found at all in 47 cases of seminoma (0%), and the positive incidence was very low in embryonal carcinoma (6.6%), although considerably higher incidences were obtained with choriocarcinoma (25.0%), yolk sac tumor (22.2%), and teratocarcinoma (13.3%). The presence and molecular species of the GM2 antigens in these human germ cell tumors were also ascertained chemically by the thin-layer chromatography (TLC) immunostaining of the ganglioside fractions prepared from primary germ cell tumors. These results indicate that seminoma specifically contains N-acetyl GM2 and no N-glycolyl GM2, suggesting that N-acetyl GM2 could be a good marker for seminoma. On the other hand, non-seminomatous germ cell tumors were characterized by the presence of N-glycolyl GM2, one of the Hanganutziu-Deicher antigens (H-D antigens). Moreover, the positive occurrence of N-glycolyl GM2 correlated very well with the degree of differentiation of non-seminomatous germ cell tumors, i.e., the differentiated tumors such as yolk sac tumors, choriocarcinoma, and teratocarcinoma had a higher positive incidence of N-glycolyl GM2 type H-D antigen but a lower positive incidence of N-acetyl GM2 when compared with embryonal carcinoma, the most undifferentiated tumors among non-seminomatous germ cell tumors.

Lectin binding in tissues from hydatidiform mole, invasive mole and choriocarcinoma to concanavalin-A, wheat germ agglutinin and peanut agglutinin.

A light microscopic analysis of lectin receptors in normal placenta and trophoblastic disease was performed utilizing biotinylated Concanavalin-A (Con-A), wheat germ agglutinin (WGA), and peanut agglutinin (PNA), in conjunction with an avidin-biotin peroxidase complex. Hydatidiform mole, invasive mole and choriocarcinoma exhibited increased receptors to Con-A and WGA compared to normal placenta. Increased reactivity to Con-A and WGA was associated merely with increased growth and proliferation of trophoblasts rather than a malignant transformation. Normal placenta, partial and complete mole generally showed moderate to strong binding with PNA after neuraminidase treatment, while invasive mole and choriocarcinoma (11 of 15 cases) generally showed minimal to absent reaction with PNA. Heterogeneity of PNA binding in choriocarcinoma was manifested by the presence of PNA reactivity in the trophoblast membrane in 2 cases wherein no prior neuraminidase treatment was given. This suggests that in some malignant trophoblasts, there is absence of sialic acid in the terminal cell surface carbohydrate groups resulting in the exposure of N-acetylgalactoseamine.

[A case of primary gastric choriocarcinoma].

A 66-year-old man with a primary gastric choriocarcinoma is presented. The pre-operative diagnosis of the gastric barium examination and an endoscopy was an unusual gastric carcinoma in the antrum. At laparotomy, an abscess in the lesser sac that had developed by a tumoral penetrance was found. Thus a total gastrectomy and a lymphadenectomy with a reconstruction was performed. The resected specimen was found to be a Borrmann 1 type tumor, and a histological examination showed it to be a choriocarcinoma with a syncytiotrophoblast, that was immunostained by human chorionic gonadotropin (HCG). The physical findings however, disclosed no tumor in the testis. The serum HCG was found to be 1,380 IU/l on the 7th postoperative day, then a pulmonary metastases appeared and progressed, and the patient died on the 22nd postoperative day.

[Trace elements in the serum and cancerous tissue in patients with trophoblastic carcinoma].

In the serum and cancer tissue of 155 patients with trophoblastic tumor and in 114 patients with benign or malignant gynecologic tumor, 22 elements (17 trace and 5 major elements) were determined by neutron activation analysis. The serum and tissue level of Cr, the serum Zn level in patients with hydatidiform mole was significantly higher than that of the control group (P less than 0.01). The serum Se level in Patients with invasive mole and choriocarcinoma was lower (less than 1.0 mumol/l), but Cu/Zn ratio was higher (greater than 2.0) in comparison with that of the controls (P less than 0.01). With remission of the disease levels of the essential trace elements went up approaching the normal levels. There was a negative relationship between the serum Zn level and hCG content. The authors point out that the low level of Zn and Se, the high level of Cr may be related to the development of trophoblastic tumors.

[The DNA content in the cell nuclei of trophoblastic tumors]. / Soderzhanie DNK v iadrakh kletok opukholei trofoblasta.

The DNA content in cytotrophoblast (CTB) and syncytiotrophoblast (STB) cell nuclei was assayed in tissue sections of 7 hydatidiform moles (HM) and 27 choriocarcinomas (CH). The procedure involved Feulgen's reaction and scanning cytophotometry. The analysis of summarized histograms showed the DNA distribution in CTB cell nuclei, on the one hand, and that in STB, on the other, to differ significantly in both the tumors. The HM studied cases were referred to as two subtypes on the basis of such parameters as modal class value, its ploidy and degree of nuclear poly- and heteroploidy of CTB and STB. These characteristics were used to identify three patterns of CH. A pronounced modal class (2c--4c) was typical of type 1. A wider range of modal class (2c--10c or 4c--8c) was observed in type 2. Type 3 of tumor was characterized by a pronounced polyploidy with the absence of the modal class. The analysis of individual CTB and histograms showed no significant differences between HM and CH with respect to the DNA content. An increase in the share of highly polyploid cells was associated with a shorter survival of patients.

[Primary choriocarcinoma of the bladder: a case report of autopsy].

The patient was a 70-year-old male with complaint of macrohematuria at the first visit to our clinic on June 10, 1986. At that time, cystoscopy revealed a thumb sized papillary tumor and a rice sized non papillary tumor, and the biopsy specimen was pathologically diagnosed as undifferentiated carcinoma. But, he refused admission. On January 30, 1987, he came back to our clinic with complaints of dyspnea, general fatigue and weight loss. Moderate lt. gynecomastia was found and the level of serum hCG-beta was detected as high as 101 ng/ml. Excretory urogram and enhanced CT revealed a large mass in the bladder. In the seventeenth day after admission, he died of lung edema and heart failure. The findings of autopsy showed a large light greenish to light brownish tumor of 10 X 10 X 3 cm in the bladder. Distant metastases were observed in internal, common iliac and paraaortic lymph nodes, but without other distant metastasis. In histological and immunohistochemical studies, the final diagnosis is choriocarcinoma of the bladder, containing syncytiotrophoblastic giant cells with hCG-beta granules as an undifferentiated carcinoma. To our knowledge this case is the eighth described in Japan. Herein we report a new case of primary choriocarcinoma of the bladder and make a brief review of the literatures.

Placental proteins in high-grade urothelial neoplasms. An immunohistochemical study of human chorionic gonadotropin, human placental lactogen, and pregnancy-specific beta-1-glycoprotein.

We examined the presence of human chorionic gonadotropin (HCG) in 16 low-grade and 47 high-grade urothelial neoplasms, including two cases with trophoblastic-like differentiation. In HCG-positive tumors, the presence of human placental lactogen (HPL) and pregnancy-specific beta-1-glycoprotein (SP-1) also was assessed. HCG immunoreactive cells were found in nine of the 47 high-grade tumors (19%), whereas none of the low-grade tumors were positive for HCG. This hormone was predominantly detected in the most undifferentiated and pleomorphic areas; however, HCG-positive cells also were found in areas of carcinoma in situ and well-differentiated transitional cell carcinoma in two cases. The serum HCG level was increased in two of the four cases studied. HPL and SP-1 immunoreactive cells were observed in seven and five cases, respectively, and it was found that tumors positive for SP-1 also were positive for HPL. Five tumors, including the two with trophoblastic differentiation, contained the three placental proteins. The HPL and SP-1 immunostained cells were usually found in the same areas of the tumor that were positive for HCG, but there was always a lower number of HPL and SP-1 immunoreactive cells than HCG immunoreactive cells. In one case, HPL and SP-1 could be found in areas of well-differentiated transitional cell carcinoma. These findings suggest that the morphologic and functional trophoblastic differentiation in urothelial carcinomas is a progressive phenomenon evolving from transitional cell carcinomas.

[Testicular germinal tumors. Current therapeutic principles]. / Tumeurs germinales testiculaires. Principes thérapeutiques actuels.

The cure rate for germinal tumors of the testes are now very high, greater than 80%, all stages and histological types taken as a whole. This calls for a revision of therapeutic indications, in light of these results and modern staging methods. In localized stages, or tumors associated with a good prognosis, therapeutic de-escalation is in progress, in order to limit the side effects. In invasive forms or forms associated with a poor prognosis, the use of more aggressive methods, such as high dose chemotherapy, should further improve prognosis.

Comparison of Coulter volumes with radiometrically determined intracellular water volumes for cultured cells.

During methotrexate-induced differentiation of cultured human choriocarcinoma (BeWo) cells, proliferation is inhibited, morphologic and biochemical changes occur, and giant, often multinucleated, cells form. We have used the increase in cell volume as a marker of the mature syncytiotrophoblastlike phenotype. Uninduced and differentiated BeWo cells are not spherical, and theoretical considerations suggested that deviations in shape could result in significant errors in Coulter volume. To determine if the values obtained by electrical pulse sizing reflected the actual mass of BeWo cells, we have evaluated the relationship between Coulter volumes and intracellular water volumes obtained using a shape-independent estimate for eight cell types. A close correlation (r2 = 0.97) was found, indicating that cell volume changes in populations of irregularly shaped cells can be accurately measured using a Coulter instrument.

[Pure testicular choriocarcinoma]. / Coriocarcinoma testicular puro.

We present a case of pure testicle choriocarcinoma in a 13-year-old male. Its first clinical manifestation consisted in a pattern of acute dyspnea caused by multiple pulmonary metastases. Alphafetoprotein investigation in serum and tissue proved negative. We carry out an immunohistochemical study of the case and discuss its histogenesis current classification and prognostic significance.

Endothelium and thrombomodulin: expression and distribution of thrombomodulin on altered endothelium and neoplastic syncytiotrophoblast.

We investigated the expression of thrombomodulin (TM) on endothelium in some pathologic states. We used the cultured endothelial cells treated with interleukin-1 (IL-1) and propagated cells by serial subculture for extended periods of time and assessed cell-surface TM molecules. We also studied the distribution of TM on surgical specimens of chorionic diseases, angiosarcoma and on several established cell lines of human choricarcinoma. Subculture of human umbilical vein endothelial cells (HUVE) up to 2 months (approximately 16 subcultures) decreased the number of cell-surface TM molecules by approximately 20% compared to the primary culture. The number of TM molecules also decreased on HUVE treated by IL-1. The treatment of the cells with IL-1 also induced change of shape. TM was found not only normal syncytiotrophoblast but also on neoplastic syncytiotrophoblast of choriocarcinoma and hydatidiform mole. However, TM was not expressed on the three established cell lines. TM was found on various types of vascular tumors, including angiosarcoma.

Malignant and normally developing trophoblastic cells of human placenta display different characteristics defined by histochemical and biochemical mapping of endogenous lectins.

Trophoblastic cells with their strictly controlled propensity for invasive growth hereby bear limited resemblance to malignant cells. Therefore, detailed description of molecular characteristics of this cell type in comparison to histologically related tumor cells may aid in defining physiologically relevant differences. In view of the potential importance of selective protein-carbohydrate interactions in recognitive processes, labelled neoglycoproteins were employed to evaluate systematically the presence and distribution of sugar receptors in tumor cells of chorionepithelioma and in trophoblastic cells of an early stage of gestation. Control reactions in glycohistochemistry, involving prolonged incubation, pH variation and use of nucleotides excluded the possibility that glycosidases or glycosyltransferases govern the sugar-specific binding to the tissue sections. Pronounced differences were detected in the expression of receptors (lectins) for alpha- and beta-glucosides as well as for N-acetylgalactosamine and N-acetylglucosamine. Further differences were revealed by probing both types of tissue with lactosylated, fucosylated, xylosylated and sialylated carrier protein. Upon developmental maturation of the normal cells in placenta the extent of binding of the neoglycoproteins decreased. The glycohistochemical differences between malignant and normally developing trophoblastic cells were found to be reflected in the alteration of expression of certain endogenous lectins, as ascertained by affinity chromatography and gel electrophoretic analysis. Consequently, we assume that the transient presence of certain endogenous lectins during early stages of gestation and their differential expression in relation to tumor cells of chorionepithelioma may be relevant for the special invasive and adhesive behavior of human trophoblastic cells.

The gonadotropin alpha-gene contains multiple protein binding domains that interact to modulate basal and cAMP-responsive transcription.

DNA sequences that modulate basal and cAMP-stimulated transcription are located within the initial 169 base pairs of the alpha-gene 5'-flanking sequence. Using DNase I protection analyses and gel-mobility shift assays, we examined in vitro the domains in the human alpha-gene 5'-flanking sequence that bind nuclear proteins extracted from JEG-3 choriocarcinoma cells. DNase I protection studies of the sequences between -236 and -100 demonstrate two major protected regions: -178 to -156 corresponding to an upstream regulatory element (URE) and -146 to -112 corresponding to 18-base pair repeated sequences that contain cAMP-responsive enhancers (CREs). Nuclear proteins extracted from JEG-3 choriocarcinoma cells bind specifically to oligonucleotides corresponding to both the URE and CRE domains as well as to a downstream domain (-99 to -72) that contains consensus CCAAT motifs on both the sense and antisense strands. Binding to a DNA fragment (-236 to -100) that contains both the URE and CRE domains was 10-fold more effective than that using either fragment alone. Binding to this multisite DNA fragment is readily disrupted using the URE sequence, but not the CRE sequence as a competitor, suggesting that the URE binding factor may stabilize DNA-protein interactions in these adjacent complexes. The amount of protein binding to each of the alpha-gene 5'-flanking domains was unaffected by treatment with 8-bromo-cAMP. These studies indicate that there are multiple adjacent protein binding domains in human alpha-gene 5'-flanking sequence that correspond to cis-acting regulatory elements including an upstream element that activates basal expression, repeated cAMP-response elements, and a downstream sequence containing consensus CCAAT box elements. Interactions between regulatory domains facilitate protein binding and synergistically stimulate alpha-gene transcription.

Structures, function, and transformational changes of the sugar chains of glycohormones.

Human chorionic gonadotropin (hCG), human luteinizing hormone, human thyroid-stimulating hormone, and human follicle-stimulating hormone are closely related family of proteins which share a common alpha-subunit. However, their sugar moieties are quite different. hCG contains five acidic asparagine-linked sugar chains. These five sugar chains are derived by sialylation from three neutral oligosaccharides: two biantennary (N-1 and N-2) and one monoantennary (N-3) complex-type oligosaccharides. Although hCG purified from the urine of pregnant women is more enriched in sialylated sugar chains than that purified from placenta, the molar ratio of N-1, N-2, and N-3 of these two hCGs are the same (1:2:1). Comparative study of the sugar moieties of the alpha- and beta-subunits of hCG revealed that alpha contains 1 mol each of N-2 and N-3, while beta contains 1 mol each of N-1 and N-2. This specific distribution of oligosaccharides at the four asparagine loci of the hCG molecule is now helping us to consider the functional role of the sugar moiety of glycohormones. hCG is produced not only by the trophoblast but also by various trophoblastic diseases. The hCGs purified from the urine of patients with hydatidiform mole contain the same oligosaccharides as normal hCG. However, those from the urine of choriocarcinoma patients contain five additional neutral oligosaccharides. In contrast, hCGs from invasive-mole patients contain three of the five oligosaccharides, specifically found in choriocarcinoma hCGs. The malignant transformational change of the sugar moiety of hCG can be explained by an increase of a fucosyltransferase, which forms the Fuc alpha 1----6GlcNAc group and by ectopic expression and subsequent modification of N-acetylglucosaminyltransferase IV. The appearance of tumor-specific sugar chains of hCG has been used to develop a new diagnostic method for invasive mole and choriocarcinoma.

Immunohistochemical studies of fetal trophoblast and maternal decidua in hydatidiform mole and choriocarcinoma.

Immunohistochemical techniques have been used to investigate the expression of fetal trophoblast antigens and the maternal leucocytic response in molar pregnancy and choriocarcinoma. The antigenic phenotype of morphologically defined trophoblast populations in complete, partial and invasive moles was analogous to that in normal pregnancy. All trophoblast phenotypes described in normal pregnancy were also identified in choriocarcinoma, suggesting that extensive differentiation into heterogeneous subgroups occurs in malignant trophoblast. The maternal leucocytic infiltrate in molar pregnancy consisted of T lymphocytes and class II MHC-positive macrophages. CD2-positive, CD3-negative lymphocytes were identified in molar decidua but not in uterine tissue in choriocarcinoma. Similarly, endometrial granulocytes were present in molar decidua but not in choriocarcinoma; these cells were associated with decidualization rather than with fetal trophoblast.