Electronic Library of Scientific Literature


Volume 47 / No. 5 / 2000


The current look at high-dose chemotherapy in breast cancer - Minireview

S. Filip, J. Petera, K. Odrážka, M. Bláha

Department of Oncology and Radiotherapy, Charles University Hospital, 500 05 Hradec Králové, Czech Republic, e-mail: filip@fnhk.cz;
Department of Hematology, Charles University Hospital, Hradec Králové, Czech Republic

High-dose chemotherapy (HDC) in high-risk breast cancer is one of the possible approaches how to improve therapeutic results, eventually, to overcome the incurability of the disease. In recent randomized studies superiority of HDC to conventional therapy has not been unambiguously established. Nevertheless, many oncologists, as well as, patients are so convinced of HDC benefits, that they are not willing to take part in randomized studies.
At an ASCO Annual Meeting (American Society of Clinical Oncology) in May 1999 in Atlanta – preliminary results of five large randomized studies phase III were presented (2 studies on metastatic breast cancer and 3 studies on high-risk breast cancer with more than 10 positive lymph nodes). The ASCO was informed of an investigation into serious scientific misconduct in a clinical trial that was presented in a plenary session of its Annual Meeting. The results of Dr. Bezwoda’s research were presented at ASCO’s Meeting as one of four plenary papers on the investigational therapy and was the only one to clearly indicate a survival benefit in the high-dose regimen.
Preliminary results presented there, however, did not confirm the original hypothesis of the high efficacy of HDC. It is necessary to wait for definite results (within two or three years, because enrollment of patients either has been finished or is being finished just now) and several parameters may change. In view of hitherto results, some investigators think that there is no need to continue in similar intensive studies. Still some believe that different modifications of therapeutic regimens or new, less toxic drugs should be tested which may lead to more effective and safer HDC.

Key words: High-dose chemotherapy, breast cancer, randomized trials.
Neoplasma, 47, 5, 2000, 261-268


Chronic myelogenous leukemia as gene activation model in oncology - Minireview

A. Zámečníková

Department of Genetics, National Cancer Institute, 833 10 Bratislava, Slovak Republic; e-mail: nci@bts.sk

Many unique features of chronic myelogenous leukemia (CML) make it as a model for studying the development of leukemia in humans. Chronic myeloid leukemia is a disease of the hematopoietic stem cell that progress in a multistep fashion. The biphasic or triphasic clinical course of the disease exemplies the multistep process of tumor progression from the indolent chronic phase to a more aggressive and terminal blast crisis. CML was the first neoplastic disease shown to be associated with consistent karyotypic abnormality now known as the Philadelphia (Ph) chromosome. The result of the Philadelphia chromosome translocation t(9;22)(q34;q11) is the transposition of the c-abl oncogene from chromosome 9 to chromosome 22, where it is fused with part of the bcr gene. The translocation generates a new hybrid bcr-abl gene which plays a crucial role in the pathogenesis of CML. Presently, CML is perhaps the best understood cancer in humans and the model of oncogenesis mediated by the Ph chromosome translocation is one of the best-characterized example of gene activation in leukemia.

Key words: Philadelphia chromosome, bcr-abl, fusion proteins, genetic testing methods.
Neoplasma, 47, 5, 2000, 269-273


Differential sensitivity of ovarian carcinoma cell lines to apoptosis induced by the IMPDH inhibitor benzamide riboside

L. Hunáková, J. Bies, J. Sedlák, J. Duraj, J. Jakubíková, X. Takácsová, L. Novotný, B. Chorváth

Cancer Research Institute, Slovak Academy of Sciences, 833 91 Bratislava, Slovak Republic, e-mail: exonhun@savba.sk;
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Kuwait University, Kuwait

The differential sensitivity of examined human ovarian carcinoma cell lines (CH1, A-2780 and SKOV-3) to the IMPDH inhibitor, benzamide riboside (BR), was demonstrated with the aid of MTT assay. Present data show that all three examined ovarian carcinoma cell lines were sensitive to the cytotoxic effects of BR in the order of sensitivity CH1, SKOV-3, A-2780, (IC50 = 2.8, 4.0 and 7.4 microM, respectively). Although the IC50 of SKOV-3 cells was similar to that previously determined by others, more than 20% of SKOV-3 cells remained viable in a plateau up to 40 microM BR concentration. This relative resistance of SKOV-3 cells to BR corresponded to the absence of BR–induced apoptosis in SKOV-3 cells, which together with clearly demonstrated sensitivity of CH1 cells to BR-induced apoptosis, established by flow cytometry (presence of nuclei with sub-G0 DNA content, Annexin V binding) and western blotting (poly-ADP-ribosyl-polymerase (PARP) cleavage), further stressed the role of drug-induced apoptosis in the overall drug-induced cytotoxicity.

Key words: Human ovarian carcinoma, cell lines, differential sensitivity, apoptosis, benzamide riboside, MTT assay, flow cytometry, poly-ADP-ribosyl-polymerase (PARP), western blotting.
Neoplasma, 47, 5, 2000, 274-279


Nucleoli in cells of the granulopoietic proliferating compartment in patients suffering from the refractory anemia of the myelodysplastic syndrome

K.Smetana, I. Jirásková, J. Čermák

Institute of Hematology and Blood Transfusion, 128 20 Prague 2, Czech Republic, e-mail: karel.smetana.@uhkt.cz

Granulocytic precursors of the granulopoietic proliferating compartment (GPC) were investigated in patients suffering from refractory anemia (RA) of myelodysplastic syndrome (MDS) to provide an information on the number of nucleoli and incidence of main nucleolar types in these cells stained with the simple cytochemical procedure for the demonstration of RNA. The results demonstrated that the incidence of main nucleolar types in all stages of the granulopoietic proliferating compartment in RA patients of MDS generally did not differ in comparison with that of control patients without a disturbed granulopoiesis. In contrast, the number of nucleoli expressed by the values of the nucleolar coefficient in all stages of GPC in RA patients was significantly smaller than in control persons. In addition, the values of the nucleolar coefficient of myeloblasts in patients with RA of MDS were close to those in patients with acute myeloid leukemias.

Key words: Granulocytic precursors, nucleoli, refractory anemia, myelodysplastic syndrome.
Neoplasma, 47, 5, 2000, 280-282


Influence of ribavirin on the micronucleus formation and in vitro proliferation of human lymphocytes

G. Joksić, M. Stanković, V. Vasić, M. Čakar, M. Jokanović

Vinča Institute of Nuclear Sciences, POB 522, 11000 Belgrade, Yugoslavia, e-mail: gjoksić@rt270.vin.bg.ac.yu;
Faculty of Pharmacy, University of Belgrade, Belgrade, Yugoslavia

Cytotoxic effects of the antiviremic drug ribavirin (1-beta-d-ribofuranosyl-1,2,4-triazole-3-carboxamide) were evaluated in vitro measuring micronucleus formation and cell proliferation kinetics in whole blood cultures employing cytokinesis block (CB) micronucleus test. The cells were exposed to ribavirin doses ranging from 0.05–0.65 micromol/ml at three different incubation times. The frequency of micronuclei in treated samples demonstrated relatively low ability of ribavirin to induce micronuclei. However, the lowest concentration of ribavirin markedly changed the frequency of mononucleated and multinucleated cells, particularly binucleated ones, which declined significantly. The decline in the frequency of binucleate cells was followed with accumulation of greater number of mononucleate cells. Decreased proliferation potential of lymphocytes treated with ribavirin indicates that cells are arrested prior to metaphase. The present investigation showed that ribavirin is capable to induce a delay in cellular proliferation at all the doses assayed. The study demonstrated that CB micronucleus assay is simple and rapid method that can be used to assess toxic effect of drugs in vitro.

Key words: Ribavirin, micronuclei, proliferation potential, human lymphocytes, toxicity.
Neoplasma, 47, 5, 2000, 283-287


The expression of proliferating cell nuclear antigen (PCNA) in leukemia cell lines HL-60 and K-562 at the light and electron microscope level

A. Grzanka, Z. Skok, A. Janiak, D.Grzanka

Department of Clinical Pathomorphology, University School of Medical Sciences, 85-094 Bydgoszcz, Poland, e-mail: grzanka@wsp.bydgoszc.pl;
Department of Biology and Environment Protection, Pedagogical University, Bydgoszcz, Poland;
Department of Clinical Pathomorphology, University School of Medical Sciences, Bydgoszcz, Poland;
University School of Medical Sciences, Bydgoszcz, Poland

PCNA antigen was localized at the light and electron microscopes level in two human leukemia cell lines HL-60 and K-562. PCNA expression was used to discriminate cycling from non-cycling cells. PCNA protein at the level of the light microscope was present in 70% of the cell in HL-60 cell line and in 65% of the cells in K-562 line. Streptavidin immuno-gold method was used for localization of PCNA expression at the ultrastructural level. Positive staining for this protein was seen as granular pattern in the nucleus and in the cytoplasm. In the nucleus the gold particles were seen to be associated with heterochromatin and euchromatin of the leukemia cells. In cytoplasm it was found on the endoplasmic reticulum and associated with ribosomes. Controls of the leukemia cells incubation with normal mouse serum showed no labelling at the light and electron microscope level.

Key words: PCNA antigen, HL-60 cell line, K-562 cell line, leukemia cell lines, ultrastructural level.
Neoplasma, 47, 5, 2000, 288-293


Bovine seminal ribonuclease induces in vitro concentration dependent apoptosis in stimulated human lymphocytes and cells from human tumor cell lines

I. Marinov, J. Souček

Institute of Hematology and Blood Transfusion, 128 20 Prague, Czech Republic; e-mail: souc@uhkt.cz

Bovine seminal ribonuclease (BS RNase), a dimeric homolog of bovine pancreatic ribonuclease has been proven to have important biological properties as aspermatogenic, antitumor, embryotoxic and immunosuppressive activities. Recently we published preliminary results concerning the ability of bovine seminal ribonuclease (BS RNase) to induce time dependent apoptosis in Con-A stimulated human lymphocytes and in human tumor cells based on DNA content and cell cycle analysis. In this study we bring more confirmative data concerning the concentration dependent in vitro induction of apoptosis in stimulated human lymphocytes and tumor cells of three human cell lines using the most sensitive and specific cytometric method for at present apoptosis determination – the indirect TUNEL. BS RNase 50 microg/ml was proven to induce 49.7, 54 and 68.1% apoptosis in the cells of the ML-2 myeloid cell line and two neuroblastoma cell lines: NB-1 and NB-2, respectively. In Con A-stimulated human lymphocytes, BS RNase also induced apoptosis, eventhough not so pronounced as in human tumor cell lines. In all cultures the induction of apoptosis was proportional to BS RNase concentration ranging from 2–50 microg/ml and correlated with proportional decrease in 3H-thymidine incorporation into the newly synthesized DNA. Side by side with the ability of BS RNase to suppress the growth of human tumors transplanted to nude mice, these biological properties determine this enzyme as a promising agent with potential clinical application.

Key words: Bovine seminal ribonuclease, apoptosis, TUNEL, flow cytometry, 3H-thymidine incorporation.
Neoplasma, 47, 5, 2000, 294-298


Prognostic significance of genetic markers in chronic myelogenous leukemia patients after bone marrow transplantation

A.Zámečníková, P.Križan, M. Mistrík

National Cancer Institute, 833 10 Bratislava, Slovak Republic, e-mail: nci@bts.sk;
Clinic of Hemathology and Transfusiology, 811 03 Bratislava, Slovak Republic

Chronic myelogenous leukemia (CML) is a malignant disease of hematopoietic stem cell with a biphasic or triphasic clinical course and most often, with a fatal outcome. Significant progress in improving outcome for patients with CML has been achieved over past years. This can be attributed to marked improvement in therapeutic protocols and increased use of bone marrow transplantation (BMT) which remains the most effective option for long–term disease control of patient with CML. The residual leukemic activity in patients after BMT remains a central clinical question. To effectively monitor minimal residual disease leukemic activity after BMT, molecular genetic techniques are currently utilized in conjunction with cytogenetic assays. Because the clinical significance of detection minimal residual disease in CML remains to be determined, we performed cytogenetic analysis and PCR amplification technique in 37 Ph+ CML patients. All patients received transplants for CML in Bratislava between years 1992 and 1999. Our results suggest that PCR positivity after transplant is of limited prognostic significance for particular individuals and can be used to identified groups of individuals at elevated risk of relapse.

Key words: Chronic myelogenous leukemia, genetic markers, bcr-abl, residual disease.
Neoplasma, 47, 5, 2000, 299-302


p53 single nucleotide polymorphisms and bladder cancer

E. Biroš, I. Kalina, J. Šalagovič, V. Habalová, M. Hrivňák, L. Valanský

Department of Medical Biology, School of Medicine, P.J. Šafárik University, 040 66 Košice, Slovak Republic, e-mail: birose@central.medic.upjs.sk;
Department of Urology, Teaching Hospital, Košice, Slovak Republic

Two p53 germline polymorphisms, a BstUI in exon 4 and a MspI in intron 6 were studied using polymerase chain reaction (PCR) based methods in 50 patients with bladder cancer and 145 healthy controls. Increased frequencies of the BstUI and MspI A2 alleles were found to be associated with statistically non-significant (p = 0.2308 and p = 0.5959) but increased odd ratios for bladder cancer (OR 1.44, 95% CI 0.82–2.27 and OR 1.20, 95% CI 0.61–2.33). Statistically significant difference between patients with bladder cancer and controls was found in the distribution of MspI genotypes. There was a significantly lower proportion of the heterozygous A1A2 genotype in all patients but not in controls (p = 0.0354). The results of this study suggest that BstUI and MspI germ line polymorphisms of the tumor suppressor gene p53 marginally modify the risk of bladder cancer.

Key words: Bladder cancer, cancer susceptibility, p53, germ line polymorphism.
Neoplasma, 47, 5, 2000, 303-306


Bone marrow and peripheral blood leptin levels in lymphoproliferative diseases – relation to the bone marrow fat and infiltration

A. Gaja, Z. Churý, L. Pecen, H. Fraňková, E. Jandáková, N. Hejlová

Masaryk Memorial Cancer Institute, 656 53 Brno, Czech Republic, e-mail: gaja@mou.cz;
Institute of Computer Science, Prague, Czech Republic

Leptin is a nonglycosylated protein produced mostly by adipocytes. The role of leptin in body weight regulation through its anorectic effect in hypothalamus is very well known. Less known are other leptin effects such as the stimulation of hematopoesis and some parts of immunity system. The role of leptin in the pathogenesis of some malignant tumors is discussed. Only a little is known about bone marrow adipocyte leptin production.
We examined leptin concentrations in the sera from peripheral blood and bone marrow, the percentage of bone marrow fat, the degree of bone marrow infiltration, the body mass index (BMI) in 42 patients with lymphoproliferative diseases. We found that bone marrow has significantly lower leptin levels (6,6 ± 10,9 ng/ml) than peripheral blood (9,1 ± 11,5 ng/ml) (p < 0.0001). Bone marrow and peripheral blood leptin levels have also a significant thin correlation (r = +0.91, p < 0.0001). Bone marrow (r = +0.55, p < 0.0005) and peripheral blood (r = +0.52, p < 0.0005) leptin concentrations are significantly correlated to BMI. Blood serum leptin (r = +0.46, p < 0.003) and bone marrow leptin (r = +0.40, p < 0.01) are related to the bone marrow fat percentage. In addition we found a negative correlation of blood serum leptin (r = -0.59, p < 0.0001) and bone marrow leptin (r = -0.42, p < 0.005) to bone marrow malignant infiltration. When we divided the patients into groups with bone marrow infiltration more than 10% and without or less than 10% infiltration, the first group had significantly lower peripheral blood (p < 0.001) and bone marrow (p < 0.02) leptin. We also confirmed a relation of bone marrow fat and infiltration (r = +0.49, p < 0.001).
Our results suggest a relationship among leptin levels in blood or bone marrow and bone marrow infiltration in lymphoproliferative diseases. This fact needs further investigation and an evaluation of its application in clinical practice.

Key words: Leptin, bone marrow fat, bone marrow infiltration, lymphoproliferative disesase.
Neoplasma, 47, 5, 2000, 307-312


Potential carcinogenic and inhibitory activity of compounds isolated from Lilium candidum L.

A. Vachálková, E. Eisenreichová, M. Haladová, P. Mučaji, B. Jóžová, L. Novotný

Cancer Research Institute, Slovak Academy of Sciences, 833 91 Bratislava, Slovak Republic, e-mail: exonvach@savba.sk;
Department of Pharmacognosy and Botany, Faculty of Pharmacy, Comenius University, Bratislava, Slovak Republic;
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Kuwait University, Kuwait

This paper deals with determination of potential carcinogenic and inhibitory activity of 10 compounds isolated from the ethanolic extract of Lilium candidum L. performed by DC polarography. The series of investigated compounds consisted of two spirostanol saponins, two pyroline derivatives, jatropham and its glucoside, flavonol kaempferol, 2-fenylethyl-alpha-l-arabinopyranosyl-(1->6)-beta-d-glucopyranoside, 2-phenylethylpalmitate and methylsuccinic acid. Carcinogenic, resp. mutagenic activity data for these compounds, with the exception of kaempferol, are not available in scientific literature.
All tested compounds were reduced in N,N-dimethylformamide in one or two well defined steps. They also formed a reversible complex with alpha-lipoic acid, a compound serving as an indicator of carcinogenic activity. The marginal diffuse current values of these complexes served as a criterion of a very low potential carcinogenicity. The inhibitory activity of isolates were studied in the presence of 12-O-tetradecanoylphorbol-13-acetate, a specific tumor promoter for an epidermal carcinogenesis, and in the presence of a polyaromatic substance 7,12-dimetylbenz(a)anthracene known as a carcinogen. The spirostanol saponins possessed the highest inhibitory activity (> 70%) and jatropham (66%). The inhibitory activity of jatropham glucoside was significantly lower (49%). Practically zero inhibitory activity was measured for the 2-phenylethylpalmitate and methylsuccinic acid.

Key words: Isolates from Lilium candidum L., carcinogenic activity, inhibitory activity, DC polarography.
Neoplasma, 47, 5, 2000, 313-318


Antiemetic efficacy of three serotonin antagonists during high-dose chemotherapy and autologous stem cell transplantation in malignant lymphoma

J. Slabý, M. Trněný, B. Procházka, P. Klener

1st Department of Medicine, General University Hospital, Charles University, 12 808 Prague 2, Czech Republic; e-mail jslaby@mbox.cesnet.cz

The aim of this study was to compare antiemetic efficacy of three serotonin antagonists, granisetron, tropisetron and ondansetron, during conditioning for autologous stem cell transplantation (ASCT).
Forty-five malignant lymphoma patients (mean age 38 years, M:F 30:15), undergoing the highly emetogenic regimen BEAM prior to ASCT, were randomized to receive IV granisetron (G) 3 mg once a day, IV tropisetron (T) 5 mg once a day, or IV ondansetron (O) 8 mg twice daily, for six days. The treatment groups were comparable with respect to age, sex and previous experience of nausea and/or vomiting. Nausea and/or emesis control failure was defined as a nausea lasting >= 4 hours and/or >= 3 episodes of vomiting/24 h, emesis control failure as >= 3 episodes of vomiting/24 h. Both the period of chemotherapy (6 days) and the whole period of observation (10 days) were evaluated.
Nausea and/or emesis control failure occurred in 24% of patients during the period of chemotherapy and in 51% of patients throughout the whole period of observation, while emesis control failed in 2% and 27% of patients, respectively. The efficacy of three serotonin antagonists was comparable during the chemotherapy period (5 patients with nausea and/or emesis control failure in the granisetron group, 2 in the tropisetron group and 4 in the ondansetron group, p = 0.40). When evaluating the whole period of observation, the antiemetic response to G and T was significantly better than to O, nausea and/or emesis control failure having occurred in 7 (47%) patients treated with G, 5 (33%) patients treated with T, and 12 (80%) patients treated with O, p = 0.03. The results concerning emesis control failures were similar, G 4 (27%), T 1 (7%), O 7 (47%), p = 0.04. Headache was the only frequent side effect of serotonin antagonists (30% incidence).
All three serotonin antagonists sufficiently controlled nausea and vomiting during high-dose chemotherapy (BEAM) administration in 67–87% of patients. In comparison with ondansetron, both tropisetron and granisetron proved to be more effective after ASCT, when emetogenic factors other than chemotherapy alone participated.

Key words: Ondansetron, tropisetron, granisetron, 5-HT3 receptor antagonist, autologous stem cell transplantation.
Neoplasma, 47, 5, 2000, 319-322


Medullary thyroid carcinoma (MTC) – clinical and molecular aspects on the basis of own experience

L. Pomorski, J. Bartkowiak, H. Pisarek, M. Bartos, J. Narębski

Clinic of Endocrinological and General Surgery, Medical University of Lodz, 93-513 Lodz, Poland;
Department of Experimental Endocrinology and Laboratory Investigations, Medical University of Lodz, Lodz, Poland;
Department of Molecular Biology, Medical University of Lodz, Lodz, Poland

In our clinic 19 615 patients were operated over 25 years on for goiter. Malignant thyroid neoplasms were found in 1049 (5.3%) patients including 875 (83.4%) women and 174 (16.6%) men. Sixty two adult patients (42 women and 20 men were operated on for medullary thyroid carcinoma (MTC). Thyroid cancer was diagnosed in this group pre– or intraoperatively in 44 (71%) patients and postoperatively, on histologic examination, in 18 (29%) patients. These patients were reoperated. Radical operations (total thyroidectomy with regional lymph node removal) were conducted in 43 (69.3%) patients and palliative ones in 19 (30.7%) patients. After MTC surgery, MEN 2A (MTC and an adrenal tumor) were diagnosed by means of imaging techniques (USG, CT) in 6 (9.7%) patients. All adrenal tumors were unilateral. Five of these patients were operated, and pheochromocytoma was confirmed by histopathologic examination. Two years after the MTC operation, 1 women was lost to follow-up. After a year, she was admitted to hospital for severe hypertension and died of cerebral hemorrhagia. Pheochromocytoma was revealed by autopsy. All patients were treated complementarily after the MTC operation. Different combinations of teleradiotherapy, chemotherapy and substitutive doses of levothyroxine were used. Ten (23.2%) of 43 patients operated radically were reoperated 1–3 years after the first operation due to loco-regional tumor recurrence. Radical reoperations were performed in 4 patients, and palliative ones in 6.
Over a 0.5–23-year follow-up period, 26 (41.9%) patients died, including 20 of cancer, and 6 of other reasons. Four out of 36 living patients have clinical or biochemical symptoms of neoplastic disease. The follow-up period of MEN 2 patients operated on ranged from 1 to 6 years. Up to now, no tumor in the second adrenal gland has been diagnosed in any of these patients. Genetic (molecular) tests performed in 31 out of 36 living patients revealed mutations of RET gene in 4 (12.9%).

Key words: Medullary thyroid carcinoma, multiple endocrine neoplasia, MEN.
Neoplasma, 47, 5, 2000, 323-326