Properties of two epithelial cell lines derived from HPV-associated cervical and vulvar lesions.

Two epithelial cell lines were established from human papilloma virus (HPV) 18 or 16 associated tumours, characterised as poorly and well differentiated squamous cell carcinomas of the cervix uteri (EC) and the vulva (GC), respectively. The cell lines are described by their morphology, biological parameters, and immunological markers. Both cell lines have undergone approximately 35 passages in vitro. HPV16 and 18 DNA are maintained integrated into the host cell DNA. Expression of epithelial cell markers--cytokeratins K1, K10, K13, K14 and involucrin, proliferation-specific proteins, proliferating cell nuclear antigen (PCNA) and Ki67 as well as the epidermal growth factor (EGF) receptor were monitored by indirect immunofluorescence studies. The cytoplasmic and membrane-associated locations of EGF receptor molecules in EC and GC cells, respectively, suggest a differently regulated expression. Studies of the HPV18 oncogene transcription revealed marked differences of amplimers between HeLa and EC cells, such as an additional fragment, probably corresponding to a E6**--E7 splice product, and a radical shift in transcription pattern observed in various sections of the tumour tissue. Injected subcutaneously into nu/nu mice both cell lines were non-tumorigenic.

Characterization of human papillomavirus type 16 activity in separate biopsies from a carcinoma of the cervix uteri.

Human papillomavirus (HPV) 16-specific nucleic acid sequences were analysed in separate biopsies taken from a patient with a poorly differentiated squamous cell carcinoma of the uterine cervix. Biopsies were obtained from histopathologically normal epithelium adjacent to the carcinomatous epithelium, the primary carcinoma and a metastatic lymph node. Signals characterizing viral DNA and oncogene transcription were obviously differentiation dependent as shown by in situ hybridization of viral nucleic acids and immunofluorescence of epithelial differentiation specific proteins. In histologically normal parts of the epithelium viral DNA was amplified at the transition from basal to maturing cells, whereas E6/E7 genes were actively transcribed mainly in maturing epithelial cells following the basal cell layer. Some of the cells in the primary carcinoma and in the metastatic lymph node expressed involucrin at increased levels. Signals for viral DNA and HPV 16-specific E6/E7 transcripts decreased in intensity during differentiation in an inverse relationship to the observed involucrin increase in those cells. The absence of Ki67 in cells expressing large amounts of involucrin as revealed by immunostaining, support the inverse correlation between differentiation of cancer cells, HPV 16 replication and E6/E7 transcription. The changes in cytokine expression may indicate an HPV 16 associated disruption of normal cytokine expression pattern in the carcinoma.

Comparison of different in situ hybridization techniques for the detection of human papillomavirus DNA in cervical smears.

Different hybridization methods were used for detection of human papillomavirus (HPV) DNA in cervical smears. The results obtained by filter in situ hybridization (FISH) are consistent with most of the reports recently published. To overcome the unsatisfactory limitations of this method, especially the difficulties to distinguish clearly between positive and negative signals, we developed an in situ hybridization protocol using a cytospin and 35S-labelled as well as biotinylated DNA-probes. For direct comparison of different methods, the samples were obtained from two groups of patients. One group were women with reiterated Papanicolaou smears III, IV; the other were women with reiterated Pap III, IV and additional histological scoring. In all cases but one, the different methods used have shown the same results. In one case the hybridization on slides using 35S-labelled as well as biotinylated probes gave a negative result, whereas the FISH method using a 32P-labelled probe allowed to detect of HPV 16 and 18 DNA only when more than 1 x 10(6) cells were present per filter. Our data demonstrate that in situ hybridization on slides is a specific and sensitive technique, which enables a clear distinction between positive and negative results using a small number of cells and which, especially with biotinylated probes, is suitable for application in routine work.

First evaluation of the Joint Clinical Registries of the Coordinating Tumor Center of Berlin.

In the present study, we report the first analysis of the clinical management [corrected] of patients with primary ovarian cancer following the summarized data of the Joint Clinical Registries of the Coordinating Tumor Center of Berlin (Dachverband Tumorzentrum Berlin e.V.). All data were summarized for the period of 2005 to 2008 regarding age, histopathology, time of surgical intervention, follow-up and survival, based on 1124 provided data sets of patients with suspected ovarian tumours. We identified 946 patients with a diagnosis of primary ovarian cancer, mostly of advanced T3 tumour stage (63.7%), FIGO III and IV stage (40.6%) or grade II and III (91%) histology. The median age at time of diagnosis was 61 years (range 15 to 94 years). Most patients (n=414, 69.8%) underwent cytoreductive surgery within one month of diagnosis. The median follow-up period was 26 months; 241 patients died in the analyzed period. The calculated 3-year survival rate was 63.3%, although the median overall survival has not yet been reached. We detected positive correlation of tumour stage (p<0.001) and of FIGO stage (p<0.001) with survival, and these were evaluated as being prognostically significant. The implementation of institutional based clinical registries as part of the modern clinical management of patients with ovarian cancer is feasible and well accepted inside the gynaecological departments of Berlin.

[Experiences with the cancer marker CA 125 in gynecologic diseases]. / Erfahrungen mit dem Krebsmarker CA 125 bei gynäkologischen Erkrankungen.

CA 125 was determined in the sera of 94 female patients afflicted with various gynecological diseases. Benign ovarian tumors and gynecologic malignancies outside the ovaries had normal CA 125 values in most cases. Patients with ovarian carcinomas in advanced stage had high levels. 6 women, who had hepatic metastases from ovarian cancer, showed a so-called prozone effect in serum titration, that means, the CA 125 concentration at first rises to reach a peak at a dilution of 1:2 to maximally 1:20, and then reveal a normal reciprocal dilution behavior. Whether hepatic metastases as a possible source of marker production are responsible for this phenomenon should be examined in future studies.