Implantation and diagnostics of endometrial receptivity.

OBJECTIVE: Literature review of endometrial receptivity in embryo implantation and its diagnostic possibilities. DESIGN: Literature review. SETTING: Department of Obstetrics and Gynecology, University Hospital, Faculty of Medicine, Palacky University, Olomouc; Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, Olomouc. RESULTS: Endometrial tissue is very dynamic, undergoing cyclic proliferation, differentiation and cell transportation, especially of immune system cells under the influence of circulating estradiol and progesterone. Endometrial remodelling during embryo implantation is controlled by decidual cells senescence and effectivity of their immunologic destruction. Endometrial receptivity can be assessed by transcriptomic profiling of endometrial biopsy using ERA system or proteomic analysis of either endometrial secretome or cervical mucus by gel electrophoresis (DIGE) or mass spectrometry (MS). CONCLUSION: With respect to recent discoveries in endometrial physiology and molecular biology, clinical application of proteomic approaches in research of potential biomarkers of endometrial receptivity could be of interest.

[Tumor cells transfer between the patient and laboratory animal as a basic methodological approach to the study of cancerogenesis and identification of biomarkers]. / Transfer nádorových bunek mezi pacientem a laboratorním zvíretem jako základní metodický prístup ke studiu kancerogeneze a identifikace biomarkeru.

INTRODUCTION: The investigation of prognostic and predictive factors for early diagnosis of tumors, their surveillance and monitoring of the impact of therapeutic modalities using hybrid laboratory models in vitro/in vivo is an experimental approach with a significant potential. It is preconditioned by the preparation of in vivo tumor models, which may face a number of potential technical difficulties. The assessment of technical success of grafting and xenotransplantation based on the type of the tumor or cell line is important for the preparation of these models and their further use for proteomic and genomic analyses. METHODS: Surgically harvested gastrointestinal tract tumor tissue was processed or stable cancer cell lines were cultivated; the viability was assessed, and subsequently the cells were inoculated subcutaneously to SCID mice with an individual duration of tumor growth, followed by its extraction. RESULTS: We analysed 140 specimens of tumor tissue including 17 specimens of esophageal cancer (viability 13/successful inoculations 0), 13 tumors of the cardia (11/0), 39 gastric tumors (24/4), 47 pancreatic tumors (34/1) and 24 specimens of colorectal cancer (22/9). 3 specimens were excluded due to histological absence of the tumor (complete remission after neoadjuvant therapy in 2 cases of esophageal carcinoma, 1 case of chronic pancreatitis). We observed successful inoculation in 17 of 28 tumor cell lines. CONCLUSION: The probability of successful grafting to the mice model in tumors of the esophagus, stomach and pancreas is significantly lower in comparison with colorectal carcinoma and cell lines generated tumors. The success rate is enhanced upon preservation of viability of the harvested tumor tissue, which depends on the sequence of clinical and laboratory algorithms with a high level of cooperation.Key words: proteomic analysis - xenotransplantation - prognostic and predictive factors - gastrointestinal tract tumors.

Striking antitumor activity of a methinium system with incorporated quinoxaline unit obtained by spontaneous cyclization.

A novel pentamethinium salt was synthesized with an unforeseen expanded conjugated quinoxaline unit directly incorporated into a pentamethinium chain. The compound exhibited high fluorescence intensity, selective mitochondrial localization, high cytotoxicity, and selectivity toward malignant cell lines, and resulted in remarkable in vivo suppression of tumor growth in mice.

Prognostic importance of selected molecular immunohistochemical markers and DNA ploidy in endometrial cancer.

UNLABELLED: The aim of the study was the analysis of the new molecular genetic immunomarkers (p53, c-erbB-2, Ki 67, bcl-2) hormonal receptors (ER, PR) and ploidy disturbances and their relation to the most important prognostic factors for endometrial cancer. The study group consisted of 135 endometrial cancer patients. Biopsies of the tumours obtained at operations were routinely histopathologically examined. Subsequenly, the immunohistochemical tumour markers were determined. The same biopsies were examined by microdissection and flow cytometric ploidy analysis and karyotyping. The findings were compared with the most important prognostic factors for endometrial cancer, mainly with clinical stage of the disease and grade. RESULTS: High expression of p53, Ki 67, c-erbB-2 and low rate of progesterone receptors was found in the prognostically unfavourable group (G 3). Aneuploidy was found in 72% in the group of poorly differentiated endometrial cancers (G 3) in contrast to 27% in the group of G1 and G2 tumours, but this difference was not statistically significant. CONCLUSIONS: Identification of p53, Ki 67, c-erbB-2, PR and determination of DNA ploidy is a useful tool to specify a group of prognostically unfavourable patients.

[Prognostic importance of selected molecular genetic immunohistochemical markers and DNA ploidy in endometrial cancer]. / Prognostický význam vybraných molekulárne-genetických imunohistochemických markeru a DNA ploidity u karcinomu endometria.

OBJECTIVE: The aim of the study was the analysis of the new molecular genetic immunomarkers (p53, c-erbB-2, Ki 67, bcl-2) hormonal receptors (ER, PR) and ploidy disturbances and their relation to the most important prognostic factors for endometrial cancer. DESIGN: Prospective study. SETTING: Dept. of Gynaecology and Obsterics, Laboratory of Experimental Medicine, Institute of Pathology, Institute of Molecular and Transplational Medicine, Medical Faculty and University Hospital, Olomouc. METHODS: The study group consisted of 88 endometrial cancer patients. The biopsies of the tumours obtained at operations were routinely histopathologically examined. Subsequently, the immunohistochemical tumormarkers were determined. The same biopsies were examined by microdissection and flow cytometric ploidy analysis and karyotyping. The findings were compared with the most important prognostic factors for endometrial cancer, mainly with clinical stage of the disease, grade and histopathological type. RESULTS: Aneuploidy was found in 71% in the group of poorly differentiated endometrial cancers (G3) in contrast to 47% in the group of G1 and G2 tumours. High expression of p53, Ki 67, c-erbB-2 and low rate of sex hormone receptors was found in the prognostically unfavourable group (G3). CONCLUSIONS: Aneuploidy seems to be an important prognostical factor for endometrial cancer patients. Identification of p53, Ki 67, c-erbB-2, ER a PR is a useful tool to specify a group of prognostically unfavourable patients.

The radiosensitising effect of olomoucine derived synthetic cyclin-dependent kinase inhibitors.

Bohemine and roscovitine are the most important representatives of the group of compounds structurally derived from olomoucine. Biologically they function as inhibitors of cyclin-dependent kinases (CDKs), the key regulators of cell cycle, which is often disrupted in cancer cells resulting in uncontrollable proliferation. Bohemine and roscovitine have demonstrated their cytostatic and cytotoxic in vitro and also in vivo effects. Currently the phase II clinical trials for roscovitine are underway. The aim of the study was to evaluate the potential in vitro radiosensitising effect of bohemine (BOH) and roscovitine (ROS). Clonogenic survival assay and human lung adenocarcinoma cell line A549 were used. Tested schedules were: A-pretreatment, B-concomitant application and C-posttreatment. Concentrations corresponded to IC10, IC25 and IC50 for BOH/ROS (0.1-30 microM). The radiation doses were 1, 2 and 3 Gy. Flow cytometry and western blot analysis were used to characterize cell cycle distribution, BrdU incorporation and DNA repair processes. The highest in vitro radiosensitising effect of BOH/ROS was observed for Schedule A in all tested concentrations (SER(37%) 1.46-3.20). Cell cycle analysis showed an inclination towards G0/G1 delay 48 hours posttreatment and unaltered level of apoptosis. Changes in the DNA repair processes were observed - inhibition of DNA-PK kinase, inhibition of BrdU incorporation, strong and enduring induction of p21 protein and long-lasting phosphorylation of gammaH2AX(Ser139). Certain low concentration activities of BOH/ROS in monotherapy were detected, mainly the activation of DNA-PK kinase. The results demonstrated strong in vitro radiosensitising effect of BOH/ROS that is concentration and especially schedule dependent. The strong cytostatic effect of the pretreatment schedule is mediated through the inhibition/rearrangements of DNA repair processes.

Prostate cancer and the food supplement, PC-SPES. Minireview.

Cancer of the prostate has now become the most frequently diagnosed cancer in the male population in America and indeed its incidence has increased in many westernised countries. Surgery, radiotherapy and androgen ablation form the mainstay of its treatment. However, these treatment modalities are ineffective in a large percentage of patients due to grade of tumor at presentation, sensitivity to radiotherapy, hormone-insensitive profile of tumor cell population, etc. Because of the woeful ineffectiveness of these standard treatment modalities on the overall survival rate from prostate cancer in the past fifty years, a more radical approach to the treatment of this cancer is justified. To this end some of the latest innovative approaches are now being tested as tools in prostate cancer research, including those pioneered in molecular and cellular biology and immunology. There has also arisen a considerable interest in the modification of diet and the use of herbs and a great interest has been stimulated in PC-SPES, a herbal food supplement sold in the USA. This product, which is composed of eight herbs, is recommended as a food supplement for those suffering from prostate cancer and there are many anecdotal, scientific and clinical claims for its efficacy. This is a review of some of the in vivo and in vitro research carried out on this product.

In vitro chemoresistance profile and expression/function of MDR associated proteins in resistant cell lines derived from CCRF-CEM, K562, A549 and MDA MB 231 parental cells.

Although cellular experiments have elucidated a number of active principles in the study of the multidrug resistance (MDR) phenomena, most of the drug resistant tumor cells were derived from different parental cell lines. This fact limits generalization of some experimental data and conclusions, and therefore we selected and characterized cell lines resistant to various anti-cancer agents derived from four parental cell lines: CEM (human T-lymphoblastic leukemia), K562 (human myeloid leukemia), A549 (human lung adenocarcinoma) and MDAMB 231 (human breast adenocarcinoma). In total we obtained a set of 42 resistant sublines, which is an excellent tool for the future studies of different aspects of MDR. In this study we report on some basic characteristics of these sublines, namely, cross-resistance to other anti-cancer drugs investigated by in vitro MTT assay, expression of MDR associated proteins (Pgp, MRP1, LRP, GST-pi and Topo IIalpha) as well as the functional activity of Pgp and MRP.

[Tumor biopsy--a trivial matter or a basic procedure?]. / Biopsie nádoru--banalita, nebo zásadní výkon?

The surgeon is frequently faced with the task of bioptic sampling of tissues from patients with tumourous diseases. The importance of such frequently minor procedures is incorrectly underrated. The importance of biopsy is in the foreground in particular at present at a time of individualization of treatment of malignant tumours, based on the development of new diagnostic-therapeutic methods. The bioptic specimen is the fundamental link in the basic decision--benignity or malignity--and also a unique source for assessment of prognostic and predictive factors on the basis of which we select the optimal therapeutic procedure. Contrary to current histopathological examination where the principles of correct collection of tissues specimens are generally known, in recent years the importance of molecular examinations is increasing where the surgeon must respect certain different principles to make the sampling successful. The authors working in a department of surgical oncology along with authors from specialized laboratories formulate rules of correct implementation of biopsies and transport of biological material in conjunction with recent laboratory methods, and based on examples of their own practice, they demonstrate how the initial approach of the surgeon can influence in a decisive way the correct diagnosis and therapeutic procedure in oncological patients.