Nonseminomatous germ cell testicular tumors clinical stage I: differentiated therapeutic approach in comparison with therapeutic approach using surveillance strategy only.

Surveillance after orchiectomy alone becomes popular for the management of clinical stage I nonseminomatous germ cell testicular tumors (CS I NSGCTT). Effort to identify patients at high risk of relapse leads to searching for risk factors of CS I NSGCTT. The aim of the study was to analyse own long-term experiences with different therapeutic approaches in CS I NSGCTT patients according to risk factors of the disease progression and to correlate these results with the group of patients who were treated with surveillance strategy only. From 11/1984 to 12/1991 a total of 145 patients with CS I NSGCTT were treated with surveillance strategy only (group A) and were followed-up to 1/2007. Patients, who had the disease progression, were treated with systemic chemotherapy. The disease progression was experienced in 52 patients (35.9 %). The overall survival rate of the patients in this group was 130/145 (89.7 %). From 1/1992 to 1/2007 a total of 323 patients with CS I NSGCTT were stratified to different risk-adapted therapeutic approaches (groups B1-3) according to histopathologic findings of primary tumor removed by inguinal orchiectomy. 111 patients (group B1) with vascular invasion and majority of embryonal carcinoma component in the primary tumor were treated with adjuvant chemotherapy (2 cycles of BEP). Disease progression developed in two patients (1.9 %). Other patients live without evidence of disease (NED). None of them died. Among 11 patients (group B2) with vascular invasion and majority with teratomatous elements in the primary tumor underwent primary retroperitoneal lymph node dissection (RPLND), 9 were found to be pathological stage I. The disease progression was observed in two patients (18.2 %), they died 87-122 months following orchiectomy. Two patients (18.2 %) with pathological stage II received adjuvant chemotherapy. Other 7 patients live with NED following RPLND. 201 patients (group B3) without vascular invasion have been followed after orchiectomy alone. They were kept under close surveillance, consisting of regular follow-up with tumor markers, chest x-ray and CT of the retroperitoneum. The disease progression was observed in 39 patients (19.4 %), who were treated with BEP chemotherapy. Three of them (7.7 %) died after a mean follow-up of 32.7 months following orchiectomy. The overall survival rate of all patients in group B1-3 was 98.4 %. Introduction of different therapeutic approaches in CS I NSGCTT patients according to risk factors of the disease progression might reduce the overall relapse rate of these patients from 35.9 % (group A) to 19.4 % (group B3) (P< 0.001). Surveillance procedure is recommended only in patients without vascular invasion in the primary tumor.

Concanavalin A receptors on the surfaces of human breast cancer cells in organ culture.

The binding of concanavalin A (Con A) at the free apical membranes of surface tumor cells in human breast cancer explants grown in organ culture was studied cytochemically with horseradish peroxidase (HRP) as a marker. On the cell membranes in aldehyde-fixed explants or explants exposed to Con A at 4 degrees C, a continuous label covered the entire free surface of the cell, which indicated the dispersed distribution of Con A binding sites. The binding of Con A at 20 degrees C resulted in discontinuous label of the cell surface, with gaps of unlabeleled membrane and partial endocytosis of the label. Incubation at 37 degrees C, following the binding of Con A and HRP at both temperatures, induced more extensive, incubation time-dependent discontinuities of the surface label that led to complete disappearance of the label from the surface and its eventual endocytosis. Con A was topically stablized on the surface only in those regions where two membranes or a different part of a folded membrane were in close contact. No differences were found in the binding, redistribution, and internalization of bound Con A among the various tumors studied.

Clonal variation in the production of tumor-associated alpha 2-macroglobulin in a malignant human melanoma and association with growth stimulation.

alpha 2-Macroglobulin (alpha 2-M) is known as a wide-spectrum proteinase inhibitor and to bind covalently certain growth factors. We have previously characterized tumor-associated alpha 2-M synthesized and secreted by human tumor cell lines. Of the cell lines studied, the melanoma cell line HMB-2 produced the largest amount of this glycoprotein. Immunofluorescence staining of cultured HMB-2 cells suggested that the cell population is heterogeneous with respect to alpha 2-M production. We have now isolated clones from the parental HMB-2 cells and characterized eight representative ones in detail. They varied considerably in the quantity of alpha 2-M secreted, from 4.2 to 46.5% of total protein. No relationship between the production of alpha 2-M by these clones and their pigmentation or tumorigenicity in nude mice was found. However, statistically there was a strong correlation between the modal chromosome number and population doubling time (r2 = 0.88, P less than 0.001) and also between the modal chromosome number and alpha 2-M production (r2 = 0.73, P less than 0.01). The growth rate of the clones correlated with the level of alpha 2-M in culture medium (r2 = 0.69, P less than 0.01). Clones with lower alpha 2-M production had a proportionally shorter population doubling time than the clones with intermediate or high production. Northern hybridization indicated quantitative variation in the alpha 2-M mRNA expressed by the parental cells and the clones, that was comparable but not identical with the quantity of alpha 2-M in the respective culture media; both parental cells and clones expressed platelet-derived growth factor A-chain mRNAs with little difference in levels. Serum-free medium from low alpha 2-M producer clones stimulated normal stationary fibroblasts significantly more than clones producing intermediate or high amounts of alpha 2-M. alpha 2-M decreased and anti-alpha 2-M IgG increased the stimulation. These results suggest that production of tumor-associated alpha 2-M is related to both autocrine and paracrine growth-stimulating activity of the tumor cells.

Cystic dysplasia of the rete testis. Case report.

Cystic dysplasia of the rete testis (CDRT) is a very rare cause of a paediatric scrotal mass often associated with renal and other genitourinary tract anomalies. These complex malformations are probably due to a developmental defect of the mesometanephric system during embryogenesis. A case of asymptomatic scrotal swelling in a 4-year-old boy is presented. Ultrasonography, showed a cystic lesion of the left testis associated with absence of the left kidney. Orchiectomy was performed because of extensive gonad involvement. Pathologic examination revealed multiple, anastomosing, irregular cystic spaces of varying sizes and shapes predominantly located in the region of the rete testis. The cysts had spread irregularly, displacing the testicular parenchyma, which was subsequently compressed under the tunica albuginea. Preoperative diagnosis of CDRT is easy if age, precise localisation, characteristic ultrasonographic features and other genitourinary malformations are considered. Other paediatric cystic lesions should be included in the differential diagnosis. It is possible to cure CDRT by orchiectomy or by conservative treatment. Nowadays the later option is preferred, but diagnosis of CDRT must be precisely established and followed by careful monitoring.

Spermatocytic seminoma associated with rhabdomyosarcoma.

A case of spermatocytic seminoma intimately associated with rhabdomyosarcoma is reported. The patient, a 51-year-old man, presented with a two-year history of right-sided testicular enlargement. Orchiectomy was performed, and a large testicular tumor was excised. Further investigations during hospitalization revealed lung, liver, and retroperitoneal lymph node metastases. Further therapy was refused, and the patient died at home two months after orchiectomy. Autopsy was not permitted. Although the great majority of spermatocytic seminomas occur in pure form, do not metastasize, and have very good prognosis, in addition to the present case, seven cases of spermatocytic seminoma associated either with rhabdomyosarcoma or undifferentiated sarcoma have been reported. Presence of the sarcomatous element is associated with aggressive behavior, metastatic disease, and very poor prognosis. It is considered that the sarcomatous element develops from the spermatocytic seminoma by anaplastic transformation.

Flow cytometry analysis of ploidy and proliferation activity in classical and spermatocytic seminoma.

In a series of 49 cases of seminomas, namely 19 classical seminomas, 21 seminomas with syncytiotrophoblastic cells and 9 spermatocytic seminomas, DNA ploidy and S-phase cell fraction of the cell cycle were estimated in paraffin-embedded histopathological material. DNA aneuploidy was detected in 16/19 classical seminomas (84%), in all seminomas with syncytiotrophoblastic cells (100%) and in 6/9 spermatocytic seminomas (67%). In three cases two distinct aneuploid stemlines were detected, in four cases regional variations in ploidy level were observed, clearly proving cellular heterogeneity within the studied specimens. No significant differences in distribution of ploidy levels of aneuploid tumors were detected either between distinct groups of seminomas or in relation to the age of the patients. On the other hand, mean values of S-phase cell fractions in our material offer statistically highly significant differences between defined groups of tumors. Spermatocytic seminomas had the highest level of proliferation activity, which is in contrast with the clinicopathological observations (relatively slow growth, rare occurrence of metastases, local malignancy). The results of proliferation activity analysis and the relatively highest incidence of diploid tumors support the theory of different origin of spermatocytic seminomas in comparison with other germ cell tumors.

Bilateral germ cell tumors of the testis.

In a retrospective study of 530 patients with testicular germ cell tumors treated between 1977 and 1993, a group of 12 patients (2.26%) with bilateral testicular tumors was analyzed. While bilateral tumors were simultaneously present in two cases (both with different histologic types), consecutive development of a tumor in the contralateral testis was observed in 10 patients 5.25 years (range, 3-13.5 years) after orchiectomy for the first tumor. The authors highlight the variability of histologic types in both testes, the need for an individual therapeutic approach with a view to previous therapy for the first tumor, the need for hormonal replacement as well as the possibility of testicular prosthesis implantation following bilateral orchiectomy.

Neo-adjuvant chemotherapy with delayed orchiectomy in patients with advanced germ cell testicular cancer.

A total of 13 patients with advanced germ cell testicular cancer underwent initial PVB chemotherapy without previous orchiectomy. Complete response (CR) of metastases was observed in 5 patients following chemotherapy alone. The residual mass persisted in 8 patients (in 5 of them in the retroperitoneum, in two patients in the lungs only and in one patient in both localizations). The residual masses were removed surgically. There were no viable malignant tumors in the removed tissue on histological examination. Delayed orchiectomy was performed simultaneously with surgical removal of the residual mass in the retroperitoneum or in the lungs in 8 patients, and in 5 patients as a separate procedure in complete responders following chemotherapy alone. Residual viable tumor in the testis was found in three patients, necrotic or fibrotic tissue in 5 patients, and mature teratoma in 5 patients. In patients with advanced germ cell testicular cancer preference must be given to early beginning of intensive chemotherapy without tissue diagnosis of primary tumor by orchiectomy. Benefit of this therapeutic approach is the timely management of acute abdominal and/or pulmonary symptoms of life-threatening distant metastases.

Monitoring of patients with non-seminomatous germ cell tumors of the testis by determination of alpha-fetoprotein and beta-human chorionic gonadotropin levels and by computed tomography.

The results of a 7-year monitoring of 230 patients with non-seminomatous testicular tumors are reported with respect to the employment of radioimmunoanalysis of alpha-fetoprotein and beta-human chorionic gonadotropin levels and CT examinations of retroperitoneum and lungs. Prior to orchiectomy, elevated levels of at least one of these markers were found in 79% of patients. After orchiectomy, tumor marker levels were in 70.4% of patients in agreement with the results of CT examinations. After the completion of chemotherapy, in more than a half of patients normal tumor marker levels and positive CT findings were observed. These results were most often due to the presence of mature teratoma. In Stage I patients the advantages of tumor marker determinations and CT examinations in the early detection of tumor progression have fully been confirmed.

Familial testicular cancer and developmental anomalies.

Familial occurrence belongs to factors followed in etiology and pathogenesis of testicular germ-cell tumors. Association with abnormal testicular development, or with other risk factors is relatively frequent. In our material 650 patients had been treated for testicular cancer in the period of 1981-1995. Familial occurrence was observed 7-times (1.08%), most frequently in combination with cryptorchidism. Individual families were analyzed in details, including HLA typing. On basis of the observations the supplementation of initial examination of each patient with suspicious testicular cancer with detailed familial history aimed also at the occurrence of urogenital developmental anomalies and tumors has been recommended. The knowledge about familial tumor occurrence in the first-degree relatives in combination with thorough testicular self-examination is being considered of great importance in the secondary prevention.

Conditions for hormone-stimulated expression of endogenous C57Bl strain-associated mammary tumor virus genome.

Full MMTV-Y gene expression can occur in dexamethasone-insulin-prolactin-stimulated cell cultured derived from C57Bl/10 mammary adenocarcinomas induced by syncarcinogenic action of chemical carcinogens (dimethylbenzanthracene), and mammotropic hormones (estrogen and prolactin). The rate of the hormone-stimulated virus production, as determined by biochemical, immunological and electron microscopical methods, was comparable to the level of spontaneous MMTV production by cells of established mouse mammary cancer cultures (CCL-51 and Mm5mt/cl). On the contrary, no virus production has been detected in hormone-stimulated cultures derived from C57Bl/10 mammary tumors induced by chemical carcinogen alone (urethan).

Cell proliferation kinetics and nuclear morphology in endometrial cancer under progesteron treatment.

Cell proliferation kinetics by using the double labeling with 3HTdR--14CTdR, and nuclear morphology were studied in 20 patients with endometrial cancer who were given progesteron as a preliminary therapeutic measure. Results of these studies indicate that we have to face the fact that in all histological types of endometrial cancer considerable variations in the amount of tumor cells undergoing secretory conversion occur due to the primary heterogeneity of the tumor cell population.

Organization of proviral DNA and protein composition of hormonally stimulated C57Bl/10 strain-associated mouse mammary tumor virus.

Two continuous lines, BL-MaTU/A1 and BL-MaTU/s6, were established from C57Bl/10 mammary adenocarcinomas induced by DMBA-prolactin-estradiol treatment. Under in vivo stimulation with dexamethasone, insulin, prolactin, and prostaglandin A1, the cells produce detectable amounts of B-type particles with biochemical properties similar to the GR-MuMTV. Analysis of restriction endonuclease-generated fragments of cellular DNA revealed identical patterns in the integration sites and internal recognition sites of MuMTV proviral equivalents in the tumor cells and normal organs of C57Bl/10 strain mice. The restriction DNA fragments of C57Bl/10-associated MuMTV proviral DNA are closely related to those of the Balb/c-associated MuMTV. These results indicate the endogenous origin of MuMTV produced in the hormonally stimulated cultures of DMBA-prolactin-estradiol-induced C57Bl/10 mammary tumors.

Expression of the alpha 2-macroglobulin receptor on human neoplastic fibroblastoid cells.

The alpha 2-macroglobulin membrane-associated receptor (alpha 2MR) has been previously detected on hepatocytes, fibroblasts, macrophages, syncytiotrophoblasts and recently on human malignant blood cells of myelomonocytic leukemia. In cells growing in vitro from human germ cell tumors alpha 2MR mRNA was detected by Northern blotting. Endocytosis of alpha 2M from culture medium was detected in these cells by indirect immunofluorescence. In cell extracts alpha 2M and its degradation products were detected by immunoblotting. The cells expressing alpha 2MR and internalizing alpha 2M were identified as fibroblasts both by their morphology and expression of vimentin intermediate filaments. The role and function of alpha 2MR receptor in the analyzed neoplastic cells of teratomatous origin is discussed.

Purification and protein composition of endogenous rat viruses.

Endogenous retroviruses are not in the majority of cases the cause of any neoplasia, except for the laboratory conditions. As far as they might serve for the evolution of pathogenic retroviruses more attention should have been paid to them. In this paper we introduce some approaches to the purification of rat endogenous retroviruses to such a degree of purity that enabled satisfactory SDS-PAGE analysis of its structural proteins. Purities of samples obtained by usual purification methods, long-term isopycnic centrifugation at a high gravity force and velocity centrifugation are compared. Protein profile of rat endogenous virus in SDS-PAGE is compared with the ones of other retroviruses. For the first time the evidence was obtained for the striking similarity between electrophoretic protein profile of rat endogenous virus WERC and feline leukemia virus. The major structural proteins of rat endogenous retrovirus and feline leukemia virus cannot be distinguished even when resolution long gradient PAGE had been employed. The accordance of electrophoretic mobilities of major structural proteins in SDS-PAGE can indicate the relatedness of retroviruses.

An in vitro study of the oncogenic effects of two variants of avian sarcoma virus B77 on rat embryonal fibroblasts.

Two variants of avian sarcoma virus B77 have been tested for their heteroinductive oncogenic in vitro effects on rat embryonal cells. Variant 22-B77V belonging to subgroup B, had been cultured for a long time exclusively on its original host, i.e. on chicks, before being used in the study. This variant of B77V yielded negative results in repeated experiments on rat cells. Variant 55-B77V belonging to subgroup C and which had been made to pass through rat cells, caused a malignant transformation in the latter. The resultant type of this interaction were virus-producing tumorous cells (designated LWF B55). This study present some of the basic characteristics of these cells.

Retrovirus-like particles produced by human embryonal cells and cell lines derived from human malignancies.

Very small amounts of retrovirus-like particles were obtained from tissue culture fluids of various human cell lines. The particles were found in almost all cultures of rapidly growing human cells (embryo fibroblasts, various types of leukemias, melanoma, urinary bladder, lung and mammary carcinomas). Morphology and some biochemical characteristics of purified samples of these particles are presented. The particles resemble C-type mammalian retroviruses.

Occurrence of carcinoembryonic antigen in tumor tissue and serum of breast cancer patients.

In 116 breast cancer patients, the levels of carcinoembryonic antigen (CEA) were determined before operation in the serum using RIA, and after operation in sections of breast tumor tissue using the immunohistological PAP technique. CEA circulating in the serum was found in 49 patients (42%). Elevated values (over 10 micrograms/l) were found in only 12 patients (10%). In histological specimens CEA positivity was found in 94 tumors (81%), however, in a majority of them the number of positive cells per section was low (1-10%). A comparison of positive and negative findings both in the serum and in the tumor specimens of individual patients showed that both serum and tumor sections were CEA positive in 40 patients (35%) and both localizations were CEA negative in 13 patients (12%). Although most patients had positive histological sections but negative sera (46%). Only 7% of patients had negative sections and positive sera. In 41 patients CEA could be examined both qualitatively (immunohistologically), and quantitatively in the cytosol of the same homogenized tumor. Of them, 30 patients (72%) had in the cytosol a CEA concentration exceeding 5 micrograms/g proteins, in 11 of the 41 patients (28%) no CEA was found. Immunohistological examination of CEA in this group gave positive results in 35 out of the 41 patients (85%), and only 6 tumors (15%) were completely negative. CEA was shown to be present in each histological type of the tumors studied, invasive ductal tumors being slightly more frequent and more positive than the lobular ones. No relation was observed to the structure of the tumors, nor to the degree of their differentiation. Thus, the examination of CEA levels can hardly contribute to the improvement of histological classification.

Biological and immunological properties of the HMB-2 human melanoma cell line.

The HMB-2 human melanoma cell line derived from a lymph node metastatis is described. After long-term cultivation in vitro the cells retained their morphology, high growth rate, xenotransplantability into immunosuppressed mice and genotypic and phenotypic markers of human melanoma cells. Upon infection of the HMB-2 cells with temperature-sensitive mutant vesicular stomatitis virus (VSVts045) at nonpermissive temperature, a complemented virus pseudotype (VSV(HMB-2] was produced carrying assembled melanoma-associated antigens. Monoclonal antibodies prepared against HMB-2 cell membrane proteins and proteins of purified VSV(HMB-2) particles showed different reactivity with various human tumor cell lines and tissues: While the RG-12 anti-HMB-2 monoclonal antibody recognized a class II tumor-associated antigen present in melanoma and carcinoma tissues, the B-6 anti-VSV(HMB-2) antibody showed selective reactivity with melanoma cells.

Chemotherapy of testicular cancer: 10-year experience.

A total of 250 patients with germ cell testicular tumors were treated by PVB chemotherapy between 1982 and 1992. Mean age of patients was 28.9 years (range 15-52). Thirty-four patients in clinical Stage II (11 patients IIA, 13 patients IIB, and 10 patients IIC) underwent primary retroperitoneal lymphadenectomy (RPL) with subsequent chemotherapy. They were followed-up for a mean of 106.3 months (range 85-125). CR was achieved in 30 patients (88.2%). Three patients relapsed. Twenty-seven patients (79.4%) are alive with no evidence of disease (NED) after a minimum of 5 years since the start of therapy. One hundred and twenty-two patients underwent primary chemotherapy for clinical Stages IM (15 patients), IIA (31 patients, IIB (48 patients) and IIC (28 patients) with RPL in cases with residual mass in the retroperitoneum. They were followed-up for a mean of 47.7 months (range 6-122). CR was achieved in 115 patients (92.7%) (75 of them received chemotherapy alone, 40 patients achieved CR following combined cytostatic-surgical treatment). Eleven patients relapsed. One hundred and nine patients (89.3%) are alive with NED. Ninety-four patients in Stages III and IV (8 patients III, 86 patients IV) underwent primary chemotherapy with additional surgical removal of residual metastases. They were followed-up for a mean of 50.5 months (range 6-125). CR was achieved in 65 patients (69.1%) (32 of them received chemotherapy alone, 33 patients achieved CR following combined cytostatic-surgical treatment). Eleven patients relapsed. Fifty-seven patients (60.6%) are alive NED. There were 11 patients with advanced germ cell testicular cancer (Stages IIC and IV) who underwent initial PVB chemotherapy without previous orchiectomy. Delayed orchiectomy was done simultaneously with surgical removal of residual mass in the retroperitoneum or in the lungs or at completion of chemotherapy alone. The toxicity of chemotherapy was moderate. There were drug-related deaths in ten patients (4%).