Expression of phosphatase of regenerating liver-3 (PRL-3) in endometrioid cancer and lymph nodes metastases.

PURPOSE: We identify the expression of PRL-3 in primary endometrioid endometrial cancer and metastases in relation to the clinicopathological characteristics. MATERIAL/METHODS: The study involved 30 patients with type I endometrial cancer. Twelve of them were diagnosed with metastases in various localization of abdomen. The PRL-3 expression was evaluated on the basis of immunohistochemistry results by the use of monoclonal antibody anti-PRL3 clone 3B6. RESULTS: The intensity of PRL-3 expression in correlation with tumor stage was statistically significant (p = 0.024). The strongest reaction was noted in cases classified as a 1a and 1b stage defined by FIGO. The strength of PRL-3 expression is significantly associated with the degree of histological tumor grade (p = 0.035). CONCLUSIONS: The strong expression of PRL-3 in the primary tumor that was significantly correlated with the grade and clinical stage suggest that PTP4A3 participates in the process of endometrial carcinogenesis.

Circulating free DNA and p53 antibodies in plasma of patients with ovarian epithelial cancers.

BACKGROUND: This study was conducted in order to evaluate the significance of circulating free DNA (CFDNA), blood plasma p53 antibodies (p53-Ab) and mutations of KRAS gene in the prognosis of ovarian epithelial cancers. PATIENTS AND METHODS: A total of 126 patients were included in this study. KRAS mutations and CFDNA were detected by means of the PCR-restriction fragment length polymorphism (PCR-RFLP) and enriched by the PCR-RFLP method. Enzyme-linked immunosorbent assay was used to analyze plasma p53-Ab. RESULTS: KRAS mutations were detected in 27 (21.4%) of examined tumors. The frequency of KRAS mutations was especially high in mucinous cancers (P < 0.001). CFDNA and p53-Ab were frequently detected in patients with serous cancers in high grade (P < 0.001). The overall survival rate was significantly lower for patients with serous tumors and CFDNA and p53-Ab-positive than negative tumors (P = 0.022 and P < 0.001, respectively). In mucinous ovarian cancer, a worse overall survival was correlated with the KRAS mutations (P = 0.03). CONCLUSIONS: The results of the present study suggested that a presence of KRAS mutations in mucinous ovarian cancer and CFDNA and p53-Ab in serous tumors was correlated with the highest risk of cancer progression.

Analysis of quality of life of women in menopause period in Poland, Greece, Belarus and Belgium using MRS Scale. A multicenter study.

PURPOSE: The aim of this study was to compare the climacteric symptoms, the activity and quality of life of women in the menopausal period from Poland, Greece, Belarus and Belgium using a Menopause Rating Scale (MRS). MATERIAL AND METHODS: The study was conducted among women over 40 years of age, from Poland (241), Belorussia (119), Greece (100), and Belgium (79). For the purpose of this research, the Polish, Russian, Belgian and Greek versions of the MRS were used. The MRS Scoring scheme is simple, i.e. the score increases point by point with the increasing severity of subjectively perceived symptoms for each of the 11 items (severity 0-no complaints, 4 scoring points-severe). The respondent provides her perception by checking one of 5 possible boxes of "severity" for each of the items. RESULTS: Mild or no complaints were reported to a similar extent by all women from these four countries. The intensity of the psychological symptoms was similar for the studied countries and did not differ significantly (P = 0.1531). Similar results we found in the somatic symptoms among the studied groups (P = 0.1421). A significant difference in the urogenital and sexual symptoms between Belgian and Belorussian women (P<0.001) was found. The frequency of menopausal symptoms was found to be significantly (P = 0.0381) higher among Belgian women in comparison to Belorussian ones. CONCLUSIONS: We found some differences between the menopausal complaints reported by women from Belgium, Belarus, Greece and Poland. Belgian women exhibited a more impaired quality of life due to a higher rate and severity of urogenital and sexual symptoms.

Evaluation of tumor angiogenesis and thymidine phosphorylase tissue expression in patients with endometrial cancer.

The formation of new blood vessels in endometrial cancer tissue is a main process, which leads to tumor progression, and is connected with tumor expansion and invasiveness. The aim of the study was evaluation of thymidine phosphorylase protein (TP) expression in human endometrial cancer cells by immunohistochemistry and comparison obtained data with intensity of angiogenesis process and clinicopathological factors as FIGO stage of disease and histopathologic grade. Endometrial cancer specimens were obtained from 55 postmenopausal patients (aged 52 to 74 years) underwent total abdominal hysterectomy with bilateral salpingo-oophorectomy. None of patients received preoperative pelvic irradiation. Histopathological typing and grading of the endometrial tumors (G-1, G-2, G-3) as well as myometrial invasion (<1/2, >1/2) were assessed using standard criteria, on hematoxylin-eosin sections. At the surgery, FIGO clinical stage of disease was determined. Thymidine phosphorylase overexpression was observed in 23 of 55 (41.8%) cases of endometrial cancer. Although we found no statistically significant differences in TP expression between histopathologic grades, particular FIGO stages showed a significant trend of increase TP tumor overexpression. Thymidine phosphorylase overexpression cases demonstrate higher intensity of angiogenesis in comparison to negative samples and results are statistically significant for t-test (p<0.0001). The most intensive new blood vessel formation was observed in G-2 of tumor differentiation grade (p=0.013 for ANOVA test) Mean angiogenic points density (APD) values in cases of G-1 histopathologic grade reached 135.7; values of G-2 and G-3 grades reached 213.8 and 162.8, respectively. Mean intensity of angiogenesis in the first FIGO stage of disease reached 160.0 APD, in stage II 205.6 APD, and in the third 286.9, respectively. Angiogenesis was more intensive in cases of advanced tumors - analysis of variance (ANOVA) confirmed statistically significant differences in APD values between FIGO stage groups (p=0.0007). In conclusion, thymidine phosphorylase expression correlates with increased microvessel density in endometrial cancer. The intensity of angiogenesis process increases according to FIGO stage of disease, which is connected with progressing of cancer disease. Thymidine phosphorylase can play an important role in endometrial cancer progression and could offer additional information about advance of disease.

Evaluation of angiogenesis, p-53 tissue protein expression and serum VEGF in patients with endometrial cancer.

Endometrial carcinoma occurs mostly in post-menopausal women. Classical methods of prognostication, as FIGO stage and histopathologic grade, could be improved by applying additional techniques, utilizing molecular biology and immunochemistry. p-53 tumor suppressor gene, the most commonly mutated gene in human cancers has been shown to play an important role in the biology of gynecologic carcinomas. Angiogenesis, a process of formation of new vessels, being connected to tumors progression and metastatic potential was shown to be linked with tumor suppressor genes expression. The aim of the study was to evaluate relationships between intensity of tumor angiogenesis, serum levels of Vascular Endothelial Growth Factor (VEGF) and tissue p-53 protein expression in endometrial adenocarcinoma. Angiogenic Point's Density (APD) was calculated in hot spots areas using the morphometric appliance. For detection of p53 protein in tumor samples, LSAB + Kit Alkaline Phosphatase (DAKO) was used. VEGF levels were assessed in patient's blood sampled before the operation. Overexpression of p53 protein was found in tumor tissue in 35.2% of cases and mean angiogenic points density was greater in p53 positive cases. Serum levels of VEGF were above the cut off level in 54.5% of patients, in those cases angiogenesis was also elevated. In cases of p53 overexpression, VEGF levels tended to be greater as compared with p53 negative cases. In conclusion, our study demonstrated that angiogenesis was more intensive in p53 positive cases, confirming the hypothesis of tumor suppressor-gene regulation of the process of neovascularization. Serum levels of VEGF were borderline-significantly higher in cases of p53 overexpression, they were also correlated to the angiogenesis. Joint assessment of angiogenesis and tumor suppressor genes expression may contribute to reliable evaluation of the biology of endometrial carcinoma.

P53 protein expression in oral squamous cell cancer in relation to some of its clinicopathological variables.

Oral squamous cell cancer develops through a multistep process by the accumulation of genetic and phenotypic changes. Loss of P53 tumor suppressor gene function represents the most common genetic lesion in human cancer. The significance of P53 expression for the development and progression of oral squamous cell cancer has still to be evaluated. The aim of this study was to estimate relationships between P53 protein expression and some clinicopathological variables of established or presumed prognostic value. A series of 129 oral squamous cell cancers was investgated retrospectively for expression of P53 protein by immunohistochemistry of paraffin-embedded tissue sections. The slides were stained with H+E and by immunohistochemistry with anti-human P53 antibody. Positive immunohistochemical staining for P53 protein was present in 75 (58%) oral cancer cases. There were no statistically significant correlations between oral cancer P53 expression and tumor site, grading, mitotic index, invasive margin type, as well as patients age and sex. Our results suggest that immunohistochemical overexpression of P53 is an important markerof accomplished neoplastic transformation in oral cavity lesions but it does not play a crucial role in the tumor progression.

Prognostic significance of CD34 expression in early cervical squamous cell carcinoma.

The aim of the study was to evaluate angiogenesis as an independent prognostic factor and to determine the correlation of the microvessel density (MD) with lymph node metastases and survival rate in 73 women operated because of invasive squamous cell carcinoma (SCC) of the uterine cervix at clinical stages lb and IIa (FIGO). The patients were divided into two groups: I--25 (34.4%) with survival rate <5 years and II--48 (65.6%) with survival rate >5 years. Angiogenesis was quantified in light microscope using an assay for CD34. The CD34 antibody intensely immunostained single endothelial cells as well as larger microvessels. In the study. differences were revealed by comparing the MD between both groups. The 5-year overall survival rate for patients with high MD was significantly worse than for those with low MD (p<0.003). A correlation was found between angiogenesis intensity and vascular involvement as well as the incidence of lymph node metastases. Thus, tissue expression of CD34 in SCC appears to be a significant prognostic indicator.

Expression of nucleolar organizer regions (NORs) in ovarian epithelial tumors.

AgNOR staining technique was tested in ovarian epithelial tumors to evaluate its diagnostic potential in distinguishing between borderline tumors and well-differentiated carcinomas. In our opinion, the AgNOR count appears useful for assessing differences only between borderline and well-differentiated serous ovarian tumors at stage I of FIGO clinical advancement.

Proliferative activity in endometrial hyperplasia and adenocarcinoma.

Studies on the proliferative activity of cells in endometrial hyperplasia and adenocarcinoma were performed using techniques detecting Proliferating Cell Nuclear Antigen (PCNA) and Nucleolar Organizer Regions (NORs). PCNA expression was defined as the percentage of nuclei showing reactivity in 200 cells per sample. The mean AgNOR count per cell was calculated following the analysis of at least 100 nuclei per sample at a magnification of x 400. Student-t test was used for the statistical analysis. The results obtained indicate that the evaluation of cell proliferative activity expressed by AgNOR count and PCNA index can help in the distinction between atypical hyperplasia and well-differentiated adenocarcinoma, and thus can serve as a useful pathological criterion.

Angiogenesis as a prognostic factor in invasive carcinoma of the uterine cervix.

The aim of the study was to evaluate angiogenesis as an independent prognostic factor and to determine the correlation between the angiogenic index (AI) and histologic grade of the neoplastic process in patients operated on for invasive carcinoma of the uterine cervix. Angiogenesis was assessed with immunohistochemical technique using a monoclonal antibody against human factor VIII--(F8/86 M0616, DAKO, Denmark). A positive correlation was revealed between the intensification of angiogenesis and the incidence of lymph node involvement and survival rate.

Tumour necrosis factor and cancer treatment: a historical review and perspectives.

At the end of 19th century William Coley, a New York surgeon, was the first to describe necrosis of the tumour induced by bacterial toxins. In 1975, a protein responsible for the induction of this process was identified and called tumor necrosis factor alpha (TNF-alpha). The application of recombinant DNA methodology has resulted in the production of large quantities of highly purified recombinant human TNF-alpha for both preclinical and clinical studies. Although TNF-alpha is still viewed by most investigators as a selective antitumor agent, its pleiotropic activities are admirably illustrated by numerous newly discovered functions of TNF-alpha in the host defenses, inflammation, pathogenesis, regulation of gene expression and differentiation. This literature survey discusses the discovery of TNF-alpha and its potential applications to antitumour therapy.

Tissue expression of VEGF as a prognostic factor in early cervical squamous cell carcinoma.

The purpose of this retrospective study was to determine whether the intensity of tumor angiogenesis, expressed as microvessel density (MD), is indeed an important parameter predicting lymph node metastasis and survival rate in 73 women operated on for early invasive squamous cell carcinoma of the uterine cervix in stages Ib and IIa (FIGO). Angiogenesis was quantified by light microscope (LM) using an assay for vascular endothelial growth factor (VEGF). In the study, differences were revealed by comparing the MD between both groups. The patient survival with high MD was significantly worse than for those with low MD (p<0.01). A correlation was found between MD and the incidence of lymph node metastases. Hence, quantitative analysis of MD used as the expression of VEGF in the each cervical squamous cell carcinomas could be useful as a significant prognostic indicator.

Patterns of immunohistochemical staining for p53 expression in hyperplastic endometrium and adenocarcinoma.

The p53, a tumour suppressor gene, is the most commonly mutated gene human cancer. In this study, we performed immunohistochemical investigations of the expression of p53 protein in hyperplastic endometrium and adenocarcinoma. Positive immunostaining was detected in 7 (30%) cases of invasive adenocarcinoma, 2 (12%) cases of simple hyperplasia with atypia and 2 (14%) cases of complex hyperplasia with atypia. In simple and complex hyperplasia without atypia staining was seen in occasional cells. The results suggested that endometrial hyperplasia is not always accompanied by p53 protein accumulation, hence its expression is not an early exponent of the neoplastic process.

Effect of the cytokine rhTNF-alpha on the population of mast cells in the growth of MethA fibrosarcoma--a TEM study.

The aim of the present study was the ultrastructural characteristics of mast cell (MC) involved in host antitumor responses induced by local (i.t.) administration of recombinant human tumor necrosis factor alpha (rhTNF-alpha) in the primary focus of methA fibrosarcoma. MC were involved in tumor interstitium remodeling. Numerous mitochondria, well-developed RER and Golgi apparatus, clusters of polyribosomes, considerable polymorphism of granules and differentiated lamellar structures which frequently presented myelinic forms were observed after rhTNF-alpha application. In the study numerous fibres of the fibrous tissue, richly vascularized, occurred in the peripheral and intermediate tumor zones. Cluster of MC and tumor cells were seen on the border of the necrotic foci. However, proteolytic enzymes released by MC cause interstitial lysis, ensuring the place for tumor growth, and are involved in angiogenesis. Thus, it is not clear whether MC contribute to the inhibition of tumor growth or have an adjunctive role in tumor progression.

Small cell carcinoma of the uterine cervix--an uncommon variant of cervical cancer with neuroendocrine features.

Small cell carcinoma (SCC) of the uterine cervix represents an uncommon variant of cervical cancer with an extremely aggressive biologic behavior, minimum survival chances and rapid and fatal clinical course. This retrospective study included 73 cases of patients treated for invasive squamous carcinoma of the uterine cervix at stages Ib and IIa at the Department of Gynecology in the years 1996-2000. Six patients (8%) with SCC were identified among all cases, sharing the clinical features of young age and early failure of appropriate radical treatment in the presence of apparently low stage disease. Neuroendocrine cellular characteristics were assessed by the biotin-streptavidin-peroxidase (LSAB) method using antibodies against neuron-specific enolase (NSE; DAKO), chromogranin A (CGA; DAKO) and synaptophysin (SYN; DAKO). All tumors examined were positive for NSE and/or CGA and/or SYN. Although the presence of neuroendocrine features appears to correlate with decreased survival, the number of patients is not large enough to determine statistical significance. However, the results confirm that SCC of the uterine cervix is one of the most aggressive tumors of the female genital tract.

[Serum lipid levels in hypercholesterolemic post-menopausal women treated with transdermal sequential combined hormone replacement therapy only or in combination with simvastatin]. / Osoczowe stezenie lipidów u kobiet z hipercholesterolemia w trakcie przezskórnej sekwencyjnej zlozonej hormonalnej terapii zastepczej skojarzonej z podawaniem statyn.

The aim of the study was to evaluate the effects of transdermal sequential combined hormone replacement therapy alone or combined with simvastatin on the serum lipid profile in hypercholesterolaemic post-menopausal women.

[Expression of PCNA antigen in cellular proliferative activity of cervical carcinoma]. / Ekspresja antygenu PCNA w ocenie aktywnosci proliferacyjnej komorek raka szyjki macicy.

The proliferative activity of cervical carcinoma was examined using proliferating cell nuclear antigen (PCNA) staining. Labelling index (LI) have been counted in sections of routinely processed paraffin embedded tissue blocks. Significant variation in PCNA LI were observed between study groups (p < 0.0235). The results obtained indicate that the evaluation of proliferative activity of cells expressed by high PCNA LI is assessed with worse prognosis.

[Heterogeneity of ovarian epithelial tumors in analysis of nucleolar organizer regions (NORs) expression]. / Heterogennosc nowotworów nablonkowych jajnika w analizie ekspresji bialek regionów organizatorów jaderkowych (NORS).

Nonhistone proteins associated with the nucleolar organizer regions (NORs) can be visualized by a silver-staining technique on paraffin-embedded tissues. The number of black dots (AgNORs) appearing on the nuclei are thought to reflect cell differentiation and can be used as a measurement of cellular proliferative activity. In this study we tested AgNOR staining in ovarian epithelial tumors to evaluate its diagnostic potential in distinguishing between borderline tumors and carcinomas.

The antitumor effect of intraperitoneal treatment with rhTNF-alpha muteins on Ehrlich ascites tumor growth.

The intraperitoneal (i.p.) treatment with recombinant human tumor necrosis factor-alpha (rhTNF-alpha) is one of the possible therapies for tumors that are confined to the abdominal cavity. Clinical trials aiming at the exploitation of the antitumor effects of rhTNF-alpha have been largely disappointing. In this model the activity of some rhTNF-alpha derivatives was studied. Ehrlich's ascites tumor (EAT) bearing Swiss albino male mice were treated i.p. three times a week with 10 micrograms/mice of rhTNF-alpha, mutein V or mutein VI for two weeks, starting on the 4th day after tumor inoculation. Control mice received PBS. The effect of the rhTNF-alpha derivatives on the course of EAT was evaluated basing on: total ascites volume (TAV); packed cell volume (PCV); total packed cell volume (TPCV); inhibitory growth rate (IGR); cellular population of EAT fluid; morphological EAT cell changes and mean survival time (MST). In the study mutein VI had only a slight effect on MST but significant on TAV- and TPCV-IGR (p < 0.001). In mice treated with rhTNF-alpha and mutein V the enhancement of MST (p < 0.01) was accompanied by TAV- and TPCV-IGR (p < 0.001). The number of EAT cells in ascites decreased after rhTNF-alpha and mutein V administration (p < 0.001). We conclude that treatment with high-dose of this modified molecule lacking the possibility of binding with p75R and not producing so intensified side effects is likely to find wider application in therapy and prevent the ascites growth just as rhTNF-alpha dosage.

Comparative studies on the ultrastructure of the rat lungs after intratumor treatment of Morris hepatoma with rhTNF-alpha and its muteins.

The aim of the present study was the comparative analysis of morphological changes found in the lungs of Buffalo rats in the course of Morris hepatoma 5123 after i.t. treatment with recombinant human TNF-alpha (rhTNF-alpha) and its muteins. Modification of the native TNF-alpha molecule and synthesis of mutagenized analogues can prevent undesirable symptoms observed in the case of therapeutic administration of rhTNF-alpha. TNF-alpha has been shown to interact with two distinct membrane receptors (TNF-R): p55R and p75R. Mutagenized mutein V binds selectively with p55R. Mutein VI fails to recognize either TNF-R. The cytokines were applied in a dose of 10 micrograms protein in a cycle of 8 days. The control group consisted of tumor-bearing animals which were given PBS. Ultrastructural examinations were based on transmission electron microscope (TEM). Mutein VI-receiving animals showed enhanced changes of cytotoxic nature. Severe damage to endothelial cells (necrosis inclusive) was observed. Blood vascular lumen showed accumulation of neutrophils and monocytes. Features of enhanced activity of endothelial cells were noted. Focally, within pulmonary alveoli conglomerates of fibrin and fragments of damaged cells were found, with erythrocytes, neutrophils and macrophages in their vicinity. The epithelium of pulmonary alveoli showed signs of considerable damage, including necrosis. The lumen of pulmonary capillaries in rhTNF-alpha-treated animals showed a predominance of eosinophils and monocytic cells. Features of endothelial stimulation were observed, although without a tendency to form microthrombi. Much less pronounced changes both in the lung capillary bed and in the alveolar epithelial cells were noted in the mutein V-given animals. Our findings confirm the possibility of peripheral activation of cells involved in the cytokine-induced antitumor response. Mutein V with the smallest effect on the lung tissue rebuilding seems to be a rhTNF-alpha derivative which can delimit the undesirable symptoms in the course of antitumor therapy reduced to i.t. injections.