H. Lauke

Further Practical Experiences in the Recognition and Management of Carcinoma in situ of the Testis

99 biopsies from the contralateral testis in patients with unilateral germ cell tumor were investigated using the semithin section technique. Four cases (4%) revealed a carcinoma in situ (CIS) pattern. Two patients underwent a local radiation (20 Gy), 2 patients received combination chemotherapy (cisplatin, etoposide and bleomycin; PEB). No tumor cells were found in control biopsies 4-8 months after therapy. Both biopsy specimens taken from radiated patients lacked also germ cells. In contrast, 1 patient who was treated with PEB and also another 1 presenting with a teratocarcinoma of apparently retroperitoneal origin and unilateral CIS revealed germ cells after chemotherapy. The present data suggest radiation therapy to be the first line treatment for CIS-bearing testes. In order to get informations about the distribution of CIS cells in the affected testes, one or two biopsies were additionally taken from macroscopically unsuspicious tissue surrounding various solid germ cell tumors (74 patients). 56 cases (76%) revealed CIS. Even considering the possibility of missing CIS, a screening biopsy is actually the only method for detecting early manifestations of germ cell tumors and should be performed routinely in contralateral testes of patients with germ cell tumors.

Leydig cell mitoses in human testes bearing early germ cell tumors

SummaryNumerous mitoses were noted in testicular tissue from adult men with early germ cell tumors. More than 15 Leydig cells undergoing mitosis were found in the interstitial compartment. The presence of specific crystalline intracytoplasmatic inclusions demonstrated for the first time that differentiated Leydig cells are capable of proliferation. Occasionally cells are difficult to discriminate during mitosis. To establish reference criteria, the light- and electron-microscopic features of the following mitotic cells were examined: Leydig cells, fibroblasts, perivascular cells, peritubular cells, and lymphocytes. Supplementary mitoses in germ cell tumors and in a case of Leydig cell tumor were investigated. In the literature, only single reports of mitoses in Leydig cells are available. The frequent incidence of Leydig cell mitosis in early germ cell tumors may be due to the presence of growth-promoting factors in the testicular tissue.

The differentiation status of primary gonadal germ cell tumors correlates inversely with telomerase activity and the expression level of the gene encoding the catalytic subunit of telomerase

BackgroundThe activity of the ribonucleoprotein enzyme telomerase is detectable in germ, stem and tumor cells. One major component of telomerase is human telomerase reverse transcriptase (hTERT), which encodes the catalytic subunit of telomerase. Here we investigate the correlation of telomerase activity and hTERT gene expression and the differentiation status of primary testicular germ cell tumors (TGCT).MethodsTelomerase activity (TA) was detected by a quantitative telomerase PCR ELISA, and hTERT mRNA expression was quantified by online RT-PCR in 42 primary testicular germ cell tumors. The control group consisted of benign testicular biopsies from infertile patients.ResultsHigh levels of telomerase activity and hTERT expression were detected in all examined undifferentiated TGCTs and in the benign testicular tissue specimens with germ cell content. In contrast, differentiated teratomas and testicular control tissue without germ cells (Sertoli-cell-only syndrome) showed no telomerase activity and only minimal hTERT expression.ConclusionsThese findings demonstrate an inverse relationship between the level of telomerase activity and hTERT mRNA expression and the differentiation state of germ cell tumors. Quantification of telomerase activity and hTERT mRNA expression enables a new molecular-diagnostic subclassification of germ cell tumors that describes their proliferation potential and differentiation status.

The Value of the AgNOR staining Method in Identifying Carcinoma in Situ Testis

An early and reliable diagnosis is necessary in order to have the chance of a curative therapy of Carcinoma in situ testis (Cis). Forty-six testicular biopsies were investigated to assess the value of the AgNOR staining method in comparison to placental alkaline phosphatase (PLAP) immunostaining. Both methods provided corresponding results and identical tumor cells were recognized in serial sections. The mean AgNOR counts per nucleus were 26.86 (19-52, SD: 2.68) for CIS cells, 8.18 (5-14, SD: 2.20) for spermatogonia and 12.96 (9-18, SD: 2.44) for Sertoli cells, without the counts overlapping within these three groups. Even single CIS cells are easily and reliably recognizable by their typical AgNOR pattern and their high AgNOR count per nucleus. The independent estimation of 9 testicular biopsies with the AgNOR staining method and the PLAP immunostaining correspondingly revealed 7 biopsies with CIS. Two biopsies lacked tumor cells. The AgNOR staining method can be recommended as an additional diagnostic tool in identifying CIS, because of the short and convenient staining procedure, low costs and the applicability on formalin-fixed and paraffin-embedded material.

In Vitro Survival of Human Neoplastic Germ Cells

Transformed germ cells give rise to the large majority of testicular tumours, nearly 90% according to Pugh (1976). There has been a very marked increase in the incidence of germ cell tumours during the last few decades, primarily in developed countries with Caucasian population (Adami et al., 1994). Other abnormalities of male reproductive health, such as cryptorchidism, hypospadias and oligospermia, though not as well documented, appear to have risen concurrently (for review see Toppari et al., 1996). Moreover, epidemiological observations report that the incidence of testicular cancer is strongly linked to the birth cohort rather than to the age of the patients (Moller et al., 1993; Bergstrom et al., 1996). These phenomena suggest that differentiating testicular germ cells in the fetal or perinatal period may be especially prone to the negative influence of yet unknown environmental factors (Skakkebaek et al., 1998).

Insulin-Like Growth Factor-Binding Protein (IGFBP)-5 in Human Testicular Tubules

Insulin-like growth factor-binding proteins constitute a protein family of six members (IGFBP-1 to -6), which bind insulin-like growth factor (IGF)-I and -II with high affinity and thereby regulate the mitogenic and metabolic activities of IGFs (Jones and Clemmons, 1995). Many tumours and neoplastic cell lines produce IGFBPs (Werner and LeRoith, 1996) and there is ample evidence for the presence of membrane-bound IGFBPs in tumour cell lines (Muller et al., 1996) indicating the existence of specific receptors for IGFBPs. Increased levels of IGFBPs, and in particular IGFBP-2, are found in the serum from patients with Wilms’ tumours (Zumkeller et al., 1993). The presence of IGFBPs in germ cell tumours has not yet been investigated systematically and there are no data available on intratubular tumour cells presenting the early stages of germ cell tumours (carcinoma-in-situ (CIS)).

Rapid Method to Detect CIS-Cells

Seminoma cells, like intratubular tumor cells (carcinoma in situ), are characterized by abundant cytoplasmic glycogen. Visualization of glycogen in histologic sections has been shown to be a valuable diagnostic tool for detecting tumor cells in testicular tissue provided that appropriate fixatives were used to preserve the glycogen in the cells. Tumor cells fixed with 5.5 % glutaraldehyde show virtually no loss of their highly soluble content. In combination with other structural characteristics even single tumor cells can be histologically diagnosed (Holstein et al., 1987).

Reinvestigation of Patients After Primary Therapy of Testicular Tumor

We studied 263 patients with a germ cell tumor of the testis to evaluate the fertility status and correlation to clinical parameters before initial treatment and after 1 year in the follow-up (mean intervall 397 days +/- 227). Between 1984 and 1995, 760 consecutive patients with germ cell tumor of the testis were treated at the Armed Forces Hospital in Hamburg, Department of Urology (Head of Dep.: M. Hartmann). Patients were followed monthly the first year, every 2 or 3 months in the following years over a minimum of 5 years.

AgNOR in Human Leydig Cell Tumors

There may be overlap between the microscopic features in benign and rare malignant Leydig cell tumors. Moreover, there is often considerable morphological similarity to normal Leydig cells causing difficulties in predicting which tumors will show a malignant behaviour (Kim et al., 1985).

Testicular Tumor Cells Pass through the Epididymal Ducts

The diagnosis of early testicular cancer is based on the histological evaluation of testicular biopsies. Like the germ cells, the tumor cells find their way along the tubular duct system of the male urogenital tract into the ejaculate. But only very few data have been published on the exfoliation of testicular tumor cells into the ejaculate in men suffering from early testicular cancer. The screening of ejaculate to detect early testicular cancer would appear to be a rather unsuccessful method.