Volume 49 (2003) No. 1
What We Currently Know about the Structure and Function of the p53 Homologue - p73 Protein: Facts, Hypotheses and Expectations
P. ČEŠKOVÁ1, D. VALÍK2, B. VOJTĚŠEK1...........................1
1Department of Experimental Oncology and 2Department of Laboratory Medicine, Masaryk Memorial Cancer Institute, Brno, Czech Republic
Corresponding author: Bořivoj Vojtěšek, Department of Experimental Oncology, Masaryk Memorial Cancer Institute, Žlutý kopec 7, 656 53 Brno, Czech Republic. E-mail: vojtesek@mou.cz.
No abstract available.
Full text. 1-8
Adult Stem Cells and Their Importance in Cell Therapy
S. FILIP1, J. MOKRÝ2, I. HRUŠKA3......................................9
1Department of Oncology and Radiotherapy, 2Department of Embryology and Histology, Charles University, Medical Faculty, Hradec Králové, Czech Republic
3Department of Philosophy and Social Science, University of Hradec Králové, Hradec Králové, Czech Republic
Corresponding author: Stanislav Filip, Department of Oncology and Radiotherapy, Charles University, Medical Faculty, 500 05 Hradec Králové, Czech Republic. Tel.: +420 49 538 46 18; e-mail: filip@fnhk.cz.
Abstract.
Full text. 9-14
Original Articles
Proteomic Analysis of Radiation-Induced Alterations in L929 Cells
S. SZKANDEROVÁ1, L. HERNYCHOVÁ1, I. KASALOVÁ1, V. VÁVROVÁ1, J. STULÍK1, M. ABEND2, D. VAN BEUNINGEN2.................................................15
1Institute of Radiobiology and Immunology, Purkyne Military Medical Academy, Hradec Králové, Czech Republic
2Institute of Radiobiology, Federal Armed Forces Medical Academy, Munich, Germany
Corresponding author: Sylva Szkanderová, Institute of Radiobiology and Immunology, Purkyne Military Medical Academy, Třebešská 1575, Hradec Králové 50001, Czech Republic. Tel.: +420 495 251 538; fax: +420 495 513 018; e-mail: skanderova@pmfhk.cz.
Abstract.
Full text. 15-17 18-21 22-25
Local IFN-gamma Therapy of HPV16-Associated Tumours
R. MIKYŠKOVÁ, J. BIEBLOVÁ, J. ŠÍMOVÁ, M. INDROVÁ, T. JANDLOVÁ, V. VONKA1, M. ŠMAHEL1, J. BUBENÍK, L. MENDOZA................................26
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic
1Department of Experimental Virology, Institute of Haematology and Blood Transfusion,
Prague, Czech Republic
Corresponding author: Romana Mikyšková, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 37 Prague 6.
Abstract.
Full text. 26-32
Two Dynamic Morphotypes of Sarcoma Cells, Asymmetric Stellate and Triangle with Leading Lamella, Are Related to Malignancy
E. POKORNÁ1, D. ZICHA2, A. CHALOUPKOVÁ1, E. MATOUŠKOVÁ1, P. VESELÝ1...................33
1Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague
2Cancer Research UK, London Research Institute, Lincoln's Inn Fields Laboratories
Corresponding author: Eva Pokorná, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 37, Prague 6, Czech Republic. Tel.: +420 224 310 234; +420 220 183 537; Fax: +420 224 310 955; e-mail: epa@img.cas.cz.
Abstract.
Full text. 33-39
D. HULÍNSKÁ1, K. ROUBALOVÁ2, J. SCHRAMLOVÁ1..................................40
1National Reference Laboratory on Borreliosis, Electron Microscopy, 2National Reference Laboratory on Herpesviridae, National Institute of Public Health, Prague, Czech Republic
Corresponding author: Dagmar Hulinská, National Institute of Public Health, Šrobárova 48, 100 42 Prague 10, Czech Republic.
Abstract.
Full text. 40-42 43-45 46-48
For their unique properties stem cells promise to be of universal use in clinical medicine, especially in regeneration of many organs and tissues in the human body. This attractive subject receives an ever growing attention of specialists from different branches of science and, no doubt, will present one of the most studied trends in medicine in the new millenium. In this communication, the authors discuss two main types of human stem cells potentially suitable for cell-based therapy. The first are the cells obtained from embryonic tissues – embryonic stem cells, the second are the cells from adult tissues – adult stem cells. Presently, harvesting and therapeutic use of embryonic stem cells are associated with many problems both methodical and ethical. Utilization of adult stem cells in cell-based therapy is a certain solution in the current state of replacement therapy. Still, we have to be aware that this is not a compromise but one of the most prospective ways to treat a variety of serious diseases. To date, it is not yet clear which way would be more suitable and it is on us which way we choose for the benefit of millions of patients. Considering the current state of knowledge, it is impossible yet to predict which stem cells – embryonic or adult - or therapeutic approaches would yield the best results. Much research is to be done and verified in practice and, at the same time, ethical problems must be resolved.
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Original Articles
Proteomic Analysis of Radiation-Induced Alterations in L929 Cells
S. SZKANDEROVÁ, L. HERNYCHOVÁ, I. KASALOVÁ, V. VÁVROVÁ, J. STULÍK, M. ABEND, D. VAN BEUNINGEN
In this study we examined the protein expression profiles in X-irradiated L929 cells to get insight into how mammalian cells respond to radiation-induced cell damage. L929 cells were irradiated with the dose of 6 Gy and cell lysates were collected at different time intervals (20 min, 12, 24, 36, 48 and 72 h). The extracted proteins were separated by 2-DE and quantified using computerized image analysis. Proteins exhibiting significant abundance alterations when comparing irradiated to unirradiated cells were identified by mass spectrometry. Using the proteomics approach we detected 47 proteins that exhibited a significant radiation-induced increase or decrease in the course of 72 h. From this group of spots 28 proteins were identified by mass spectrometry and of these 24 proteins exhibited minimally 2-fold differences in mean abundance values in comparison to controls. The identified proteins represent diverse sets of proteins participating either in protective and reparative cell responses or in induction of apoptosis and oncogenesis. The results document that proteomics is a useful method for unravelling the molecular mechanisms involved in cell reaction to ionizing radiation.
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Local IFN-gamma Therapy of HPV16-Associated Tumours
R. MIKYŠKOVÁ, J. BIEBLOVÁ, J. ŠÍMOVÁ, M. INDROVÁ, T. JANDLOVÁ, V. VONKA, M. ŠMAHEL, J. BUBENÍK, L. MENDOZA
We have examined whether peritumoral administration of IFN-gamma can inhibit growth of HPV16-associated, MHC class I– tumour MK16/1/IIIABC (MK16) transplanted in syngeneic mice. It has been found that peritumoral administration of recombinant IFN-gamma performed on days 0-11 after tumour challenge inhibited growth of MK16 s.c. tumour transplants. If the therapy with IFN-g was started when the tumours have already reached a palpable size, the IFN-gamma administration was without any effect. To investigate the antitumour effects of IFN-gamma in a clinically more relevant setting, surgical minimal residual tumour disease was utilized. Subcutaneously growing MK16 carcinomas, 8-12 mm in diameter, were removed and the operated mice were injected with IFN-gamma on days 3-14 after the operation at the site of surgery. Treatment with IFN-gamma resulted in a moderate, reproducible, but statistically insignificant inhibition of tumour recurrences. In the next experiments we have addressed a question whether the tumour-inhibitory effect of IFN-gamma was due to the upregulation of MHC class I molecule expression on MK16 tumour cells. IFN-gamma-treated and control mice were sacrificed, their tumours were explanted, and the expression of MHC class I molecules on the MK16 tumour cells was examined. As presumed, the MHC class I expression on the cells of IFN-gamma-treated tumours, as well as on their lung metastases, was upregulated. However, an unexpected moderate upregulation of the MHC class I expression was also observed on MK16 tumours from the control, exogenous IFN-gamma uninjected mice. Cytofluorometric analysis of the in vivo transplanted MK16 tumours from both groups has excluded that the increased percentage of the MHC class I molecules on the tumour cell populations could be due to the infiltration of the tumours with MHC class I+ leukocytes, since no expression of MHC class II, CD11b, CD80/CD86, and CD11c molecules in the MK16 cell population was observed.
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Two Dynamic Morphotypes of Sarcoma Cells, Asymmetric Stellate and Triangle with Leading Lamella, Are Related to Malignancy
E. POKORNÁ, D. ZICHA, A. CHALOUPKOVÁ, E. MATOUŠKOVÁ, P. VESELÝ
A notion of the dynamic morphotype was developed as a conjunction between cell shape and migration. This enabled the investigation of the relationship between malignancy and patterns of dynamic morphology in neoplastic cells in vitro. Time?lapse cinemicroscopy was used to analyse the cell behaviour of three rat neoplastic cell lines (K2, T15, and A8), differing in metastatic potential, that were instrumental in revealing a coincidence between high migratory activity and appearance of the 3D structure of actin cables in high-malignant A8 cells (Pokorná et al., 1994). A set of criteria was established for visual classification of cell morphology. Matching the pattern of cell morphology with locomotory activity led to identification of four dynamic morphotypes. Cell speed was determined by tracking and the dynamic morphotypes assigned by the operator. All the three cell populations were studied for incidence of the dynamic morphotypes in culture media differing in pH: 6.6 simulating acid extracellular condition in tumours, physiological 7.4, and alkaline 8.2. The results showed that acid pH stimulated motile activity in the intermediate-malignant T15 and most malignant A8 cells. The T15 and A8 cells also manifested a prolonged continuation of fast locomotion in the early G1 phase and displayed a prevalence of two fast moving dynamic morphotypes: asymmetric stellate and triangle with leading lamella.
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Interaction of Borrelia burgdorferi Sensu Lato with Epstein-Barr Virus in Lymphoblastoid Cells
D. HULÍNSKÁ, K. ROUBALOVÁ, J. SCHRAMLOVÁ
Since the possibility of interruption of latent EBV infection has been suggested by the induction of the lytic virus cycle with chemical substances, other viruses, and by immunosuppression, we hypothesized that the same effect might happen in B. burgdorferi sensu lato infection as happens in Lyme disease patients with positive serology for both agents. We have observed EBV replication in lymphoblastoid cells after superinfection with B. garinii and B. afzelii strains after 1 and 4 h of their interaction. We found that viral and borrelial antigens persisted in the lymphoblasts for 3 and 4 days. Morphological and functional transformation of both agents facilitate their transfer to daughter cells. Association with lymphoblasts and internalization of B. garinii by tube phagocytosis increased replication of viruses more successfully than B. afzelii and chemical inductors. Demonstration of such findings must be interpreted cautiously, but may prove a mixed borrelial and viral cause of severe neurological disease.
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