Electronic Library of Scientific Literature


Volume 45 / No. 2 / 1998

The multidrug resistance in human leukemias

O. Zubercová, O. Babušíková

Cancer Research Institute, Slovak Academy of Sciences, 833 91 Bratislava, Slovakia

Leukemia/lymphoma cells, clinically refractory to therapy are often associated with expression of P-glycoprotein (P-gp), which is encoded by the multidrug resistance (MDR) gene, mdr1. Cell lines expressing mdr1 exhibit resistance to several structurally unrelated lipophilic drugs, such as anthracyclines, vinca alkaloids, and epopodophyllotoxins. This MDR can be conferred to drug- sensitive cells mdr1 cDNA transfer. In resistant cells, MDR is characterized by overexpression of P-gp and by the enhanced efflux, and P-gp fluorescence probe, rhodamine 123 (Rh 123). This can be circumvented by addition of certain non-cytotoxic drugs, such as verapamil and cyclosporin A.

Key words: MDR, human leukemia.
pp. 53-59

Prediction of lung cancer mortality in four Central European countries, 1990-2009

A. Kubík, I. Pleško, J. Reissigová

Pneumology and Thoracic Surgery Clinic, Third Medical Faculty, Charles University, 180 81 Prague, Czech Republic;
National Cancer Registry, Bratislava, Slovakia;
Cancer Research Institute of the Slovak Academy of Sciences, Bratislava, Slovakia

During the post-war decades the cancer mortality in Europe has undergone deep changes. In the 1980s, remarkable increases in lung cancer mortality in the Central and Eastern European area resulted in rates equaling or exceeding those in most Western countries. In the present work, the future development of the lung cancer epidemic has been assessed in four Central European countries (Austria, Czech Republic, Hungary, and Slovakia) for the period 1900-2009, taking into consideration previously observed lung cancer mortality trends (1960-1989), in the same countries. The estimation of the predicted mortality trends was based on log-linear Poisson regression age/period/cohort model, using GLIM for calculation. In the twenty-year period from 1985-1989 to 2005-2009, the age-adjusted (world standard) lung cancer mortality rates for men are predicted to increase in Hungary and Slovakia, and show little change in Austria and the Czech Republic. For women, approximately exponential increases in lung cancer mortality rates (both adjusted all-age, and age-specific at young adult ages up to 44 years) can be expected, with highest rates in Hungary, intermediate in the Czech Republic and Austria, and lowest in Slovakia. Lung cancer mortality in women is still much smaller than in men, however, rapidly increasing, with less variation in trends between countries than in men. The current and predicted high and/or increasing lung cancer mortality rates in the countries under study, presumed to be associated with elevated exposure to respiratory carcinogens, mainly cigarette smoke, in previous decades, underlines that the control of smoking continues to be a priority among approaches to cancer prevention.

Key words: Lung cancer, mortality, prediction, statistical model, smoking, epidemiology.
pp. 60-67

Antitumor properties of boron complexes with hydroxy biguanide and salicyl hydroxamic acid against Ehrlich ascites carcinoma

P. Ghosh, S.P. Bag, B. Sur, P. Sur

Department of Chemistry, Jadavpur University, Calcutta, 700 032, India;
Division of Pharmacology and Experimental Therapeutics, Indian Institute of Chemical Biology, Calcutta, 700 032 India

A new derivative of hydroxamic acid, hydroxy biguanido hydrochloride monohydrate and its boron derivative, dihydroxy-oxybiguanido boron (III) hydrochloride monohydrate were synthesized. Another boron compound, hydroxo-salicylhydroxamato boron (III) was synthesized from known salicyl hydroxamic acid. Antitumor properties of all the compounds evaluated against Ehrlich ascites carcinoma in mice show enhanced survival time when boron is incorporated in the compounds. Hematological parameters, alkaline phosphatase in serum of the treated animals show minimum toxic effects after boron is coupled with their respective hydroxamic acids.

Key words: Boron compounds, Ehrlich ascites carcinoma.
pp. 68-72

Effect of hyperthermia on transmembrane potential of HeLa cells - a flow cytometric analysis

P. Remani, V. Ostapenko, K. Akagi, Y. Tanaka

Division of Cancer Research, Regional Cancer Centre, 695 011 Thiruvananthapuram, India;
Department of Radiology, Kansai Medical University, Osaka, Japan

The effect of hyperthermia on transmembrane potential was studied in HeLa cells in vitro using a 3',3'-dipentyl oxacarbocyanine [Di-0-C5(3)], a lipophilic cation probe that equilibrates across the plasma membrane according to the transmembrane potential. Uptake of the fluorescent probe was measured by flow cytometry. The flourescent intensity (FI) increased with increase in temperature, and the increase was statistically significant when the duration of heat treatment was 30 minutes or more. At each temperature studied the depolarization was higher after longer duration of heat treatment (p value: 41°C < 0.05; 42°C < 0.005; 43°C < 0.001 and 44°C < 0.001, respectively). The lack of significant depolarization after shorter duration of heating, particularly at lower temperatures could be due to the repair of membrane damage that could have occurred in the holding interval between heating and measurement. The results suggest that depolarization of membrane potential, i.e. increase in the intracellular cation concentration, can be considered as an indicator of cell injury by hyperthermia and may be mechanistically related to cell death by heat treatment. The technique may be suitable for studying repair of damage after hyperthermia.

Key words: Membrane potential, hyperthermia, fluorescent dye, flow cytometry, HeLa cells.
pp. 73-76

Migration induction of contact inhibited C3H 10T1/2 cells by protein kinase C (PKC) dependent process

J. Miłoszewska, D. Kowalczyk, P. Janik

Department of Cell Biology, The Maria Skłodowska Memorial Cancer Center, 02-781 Warsaw, Poland

Treatment of confluent contact inhibited 10T1/2 cells with TPA or OAG induced a dramatic increase of the number of migrating cells, on cover slides inserted into culture dishes. When cover slides were coated with collagen IV or fibronectin, there was a similar increase of the number of migrating cells. RT PCR showed the presence of alpha PKC gene transcripts and the lack of beta and gamma PKC. Western blot analysis showed translocation of 80 kD alpha PKC to membranous fraction following brief treatment with TPA, and down-regulation of PKC after longer exposure to TPA. Collagen IV and fibronectin treatment of 10T1/2 cells induced MAP kinase, (MEK) kinase in the presence and in absence of FCS. Signal transduction pathway depending on protein kinase C and integrin receptors activation appears to facilitate migration of 10T1/2 cells and may be involved in the mechanism of the escape from contact inhibition of movement.

Key words: Migration, protein kinase C, integrin receptors.
pp. 77-80

Endothelin-1 and 5-fluorouracil-induced cardiotoxicity

C. Porta, M. Moroni, S. Ferrari, G. Nastasi

Department of Internal Medicine and Medical Therapy,
Section of Internal Medicine and Medical Oncology and
Section of Internal Medicine and Nephrology, University of Pavia, I.R.C.C.S. Policlinico San Matteo, 27100 Pavia, Italy

Cardiotoxicity is an uncommon side-effect of 5-FU-based chemotherapy. Coronary artery vasospasms have been postulated to be involved in the pathogenesis of this rare but serious problem. We found high plasma levels of ET-1, a potent natural vasoconstrictor, in two patients who experienced two of the commonest clinical manifestations of 5-FU-induced cardiac toxicity - i.e., angina pectoris and chronic heart failure. We, therefore, propose ET-1 as the ultimate mediator of this toxicity, even though the mechanism of ET-1 increase in peripheral venous blood is still unknown. Finally, another important question still remains unresolved: is the release of ET-1 from normal coronary endothelial cells the prime cause or simply the consequence of 5-FU-related cardiotoxicity?

Key words: 5-Fluorouracil, cardiotoxicity, endothelin-1.
pp. 81-82

Immunohistochemical analysis of the functional status of estrogen receptor cascade in breast cancer

Z. Kolář, J. Ehrmann Jr., M. Dušková

Institute of Pathology, Faculty of Medicine, Palacký University, 77 515 Olomouc, Czech Republic;
Centre of Molecular Biology and Medicine, Palacký University, Laboratory of Molecular Pathology, Olomouc, Czech Republic

Hormone receptor expression in human breast cancer cells does not always reflect tumor response to therapy. Thus, relations between hormone receptor status and other parameters need further examination. The aim of this study was to test the ability of estrogen receptors (ER) to induce progesterone receptor (PR) synthesis as well as to test their role in the regulation of cell proliferation. Measurement of the relation between expressions of ER and the estrogen receptor related protein p29 (ERRP) was a further goal. The results show that some hormone receptor phenotypes are closely related to tumor proliferative activity: in the ER-group, especially ER-PR-phenotype, proliferative activity shows no obvious relationship to phenotype status, suggesting that proliferation in this group probably is not under estrogen control, while in the ER+ group, PR expression was related to reduced proliferation. There was no clear association between ERRP and nuclear ER but the highest ERRP expression was most closely related to ER negative (ER-) or slightly positive (ER±) hormone receptor statuses. Tumors with these phenotypes are known to have a poorer prognosis. The conclusion drawn is that simultaneous estimation of proliferative activity, ERRP p29 expression and a comparison with ER/PR hormone receptor phenotype, could provide pathologist with a valuable tool for predicting hormone response and prognosis in breast cancer patients.

Key words: Breast cancer, estrogen receptor, function, proliferation, prognosis.
pp. 83-87

The relationship of HLA-DR, CD38 and CD71 markers to activation, proliferation and differentiation of some human leukemia and lymphoma cells

M. Glasová, E. Koníková, J. Stašáková, O. Babušíková

Cancer Research Institute, Slovak Academy of Sciences, 833 91 Bratislava, Slovakia

We investigated the expression-percentage as well as MESF values ("molecules of equivalent soluble fluorochrom" that represent approximately the density of marker expression) of HLA-DR, CD71 and CD38 markers in some human leukemias (ALL, AML, CLL, CML) and lymphomas. They are non-lineage restricted and are supposed to be activation markers except for cases where they represent pathological phenotype like HLA-DR in pre B-ALL, CD38 in some M0 AML or in plasmocytoma or CD38 and CD71 in less mature T-ALL. We used flow cytometry, immunofluorescent staining, DNA staining by propidium iodide and quantification by calibration particles.
We demonstrated increased MESF values of HLA-DR compared with controls in all investigated disorders, what could have a prognostic value. We demonstrated significantly higher MESF values of HLA-DR in cALL (37 300-46 000) in comparison with AML (9400-12 400), what could represent another important parameter when distinguishing between these two groups of leukemia. In cells of CML patients with lower CD38% and CD71% increased MESF values (5100 for CD38 and 7900 for CD71), were found while in some T-ALL, AML and cALL patients with high percentages of CD71 and CD38 there were lower MESF values what could indicate a possible connection of higher stage of cell maturation with increased density of CD38 and CD71 markers.
We investigated possible relationship between percentage of expression of HLA-DR, CD38 and CD71 and proliferation rate by DNA analysis of the cell cycle. In a group of non-Hodgkin's lymphoma patients, there was no significant increase of proliferation index of malignant cells compared with control. The correlation between percentage of expression of mentioned parameters and proliferation index was not significant. In one patient with Burkitt's lymphoma we demonstrated significant increase of proliferation index of CD71+ subpopulation compared with CD71- one, what indicates that in aggressive form of NHL CD71 can be evaluated not only as activation but also as proliferation marker.

Key words: Activation, proliferation, CD38, CD71, HLA-DR, antigen density, MESF, leukemia, lymphoma, flow cytometry.
pp. 88-95

Early changes in flow cytometric DNA profiles induced by californium-252 neutron brachytherapy in squamocellular carcinomas of the uterine cervix

T. Tačev, J. Žaloudík, L. Janáková, V. Vagunda

Masaryk Memorial Cancer Institute, 656 53 Brno, Czech Republic

Ninety-five squamocellular carcinomas of the uterine cervix, clinical Stages II and III, were treated by either four schedules combining 252-californium neutron-gamma-radiotherapy with different proportions of a neutron component (9, 6 and 3 Gy) or gamma-irradiation alone. Flow cytometric DNA profiles were obtainable in 72 cases before treatment and 56 cases were monitored for DNA content by flow cytometry (FCM) in weekly intervals by analysis of sequential microbiopsies for one month during and after radiotherapy. DNA aneuploidy was reduced from 40% (25/63) to 19% (9/47) one week within therapy in neutron-treated groups, but not after initial gamma-radiotherapy alone. Extinction of DNA aneuploid subpopulations occurred after neutron therapy in all remaining aneuploid tumors (9/9) during further monitoring, but only in 40% (2/5) of tumors after sole gamma-irradiation. In contrast, proliferation index by more than 50% was more often achieved in groups with a higher gamma-radiation component than after neutrons only. When all therapy-induced DNA flow cytometric events are taken together for evaluation of the effects of various radiotherapy schedules, it appears that the regimen with the maximal neutron dose may not be optimal for all tumors. It is hypothesized that the differences in the early flow cytometric DNA profiles may select the DNA aneuploid squamous cell uterine cervical carcinomas as candidates for combined neutron-brachytherapy, while highly proliferating DNA near-diploid tumors may profit more from treatment with a higher gamma-radiotherapy component. However, these early DNA flow cytometric findings need to be correlated with clinical course of the disease to validate this hypothesis, a process which will be completed at the end of the expected five-year clinical outcome in 2000.

Key words: Uterine cervix carcinoma, 252-Cf neutron brachytherapy, DNA flow cytometry.
pp. 96-101

Cytoplasmic liver cell inclusions - a typical feature of porphyria cutanea tarda - are absent in porphyria-related hepatic neoplasias

F. Fakan, A. Chlumská, J. Krijt, L. Kočová

Department of Pathology, Faculty of Medicine, Charles University, 305 99 Plzeň, Czech Republic;
2nd Department of Pathology, 1st Faculty of Medicine, Charles University, Prague, Czech Republic;
Department of Pathological Physiology, 1st Faculty of Medicine, Charles University, Prague, Czech Republic

Crystalline cytoplasmic needle-shaped inclusions in hepatocytes are considered to represent a specific morphological feature of porphyria cutanea tarda (PCT) and experimental PCT-like porphyrias. The cytoplasmic inclusions, however, are absent in hyperplastic hepatic nodules and hepatocellular carcinomas arising in the course of these conditions. It is assumed that porphyrins and related substances accumulated in hyperplastic and neoplastic hepatic lesions differ from those found in non-neoplastic liver tissue: the highly carboxylated porphyrins are stored in both sites, the crystal-forming substance only in non-proliferating liver tissue.

Key words: Porphyria, liver, inclusions, hyperplastic nodules, hepatocellular carcinoma.
pp. 102-106

Dose-response relationship for elective neck irradiation of head and neck cancer - facts and controversies

R. Suwinski, B. Maciejewski, H.R. Withers

Center of Oncology, M. Sklodowska-Curie Memorial Institute, Branch Gliwice, 44-101 Gliwice, Poland;
Department of Radiation Oncology, UCLA Medical Center, Los Angeles, USA

The aim of this study is to assign dose-response relationship for subclinical neck metastases of squamous cell head and neck cancer based on extensive survey of 24 data sets collected from the literature. Neck relapse rates (NRR) without and after elective (ENI) or preoperative irradiation were estimated for each site and stage of primary tumor and the reduction in neck relapse rate was calculated.
An average NRR without ENI was 22% (12-35%) and only 2.5% (0-10%) after the ENI with total dose of 46-50 Gy which gives high reduction rate in the risk of neck recurrences being on the average 89% and 42% (0-46%) after preoperative irradiation using 22-30 Gy. Dose response curve for elective and preoperative irradiation have shown that 50 Gy in 2 Gy fraction reduces the incidence of neck relapses in the N0 patients by more than 90% and only by less than 50% after total doses lower than 30 Gy. No correlation between the risk of neck metastases without ENI and the reduction in neck relapses after ENI was found.

Key words: Reduction in the risk of neck metastases, elective neck irradiation, dose-response relationship.
pp. 107-112

Immuno-ultrastructural localization of p53 protein in patients with laryngeal carcinoma

R. Sujkowska, A. Grzanka, D. Burduk

Department of Clinical Pathomorphology, University School of Medical Sciences, 85-094 Bydgoszcz, Poland;
Department of Laryngology, University School of Medical Sciences, Bydgoszcz, Poland

Protein p53 was localized ultrastructurally with 5 nm gold-streptavidin particles in cells of laryngeal carcinoma embedded in Epon 815. Postembedding technique to study p53 protein was used. Protein p53 was found predominantly in nucleus of the cells but the label was also observed in cytoplasm of the cells. Five of 15 samples from patients with laryngeal carcinoma showed positive labeling for p53, (33.3% of all examined samples). Controls of tumor cells incubation with normal mouse serum showed no labeling with gold streptavidin particles.

Key words: Laryngeal carcinoma, p53 protein, gold streptavidin method, immunoelectron microscopy.
pp. 113-116