Electronic Library of Scientific Literature


Volume 44 / No. 4 / 1997

The leukocyte common antigen (CD45) on human pre-B leukemia cells: variant glycoprotein form expression during the cell exposure to phorbol ester is blocked by a nonselective protein kinase inhibitor H7

J. Duraj, J. Sedlák, P. Rauko, B. Chorváth

Department of Molecular Immunology, Cancer Research Institute, Slovak Academy of Sciences, 812 32 Bratislava, Slovakia;
Department of Experimental Tumor Therapy, Cancer Research Institute, Slovak Academy of Sciences Bratislava, Slovakia

The human pre-B acute lymphoblastic leukemia cell line REH6 was utilized for characterization of CD45 glycoprotein by monoclonal antibodies (mAb) recognizing four distinct CD45 antigen specificities, i.e. nonrestricted CD45, restricted CD45RA, CD45RB and CD45R0. Immunoprecipitation revealed two antigen specificities on REH6 cells of m.w. 220 kDa and 190 kDa, both presenting wide range of isoelectric point pI~6.0-7.5. Nonrestricted CD45 epitopes were not affected by the sialyl acid cleavage with sodium metaperiodate or neuraminidase, but were sensitive to both, tunicamycin, the N-glycosylation inhibitor and monensin, an inhibitor of protein transport through the Golgi compartment. O-sialoglycoprotein endopeptidase from Pasteurella haemolytica A1 partially cleaved CD45RA and CD45RB epitopes, while nonrestricted CD45 determinants were not affected by this enzyme. Limited proteolysis of this antigen resulted in the appearance of 160-180 kDa peptide domains which retained CD45 epitopes. Further, the treatment of cells with phorbol myristate acetate (PMA) induced marked down-regulation of 220 and 190 kDa isoforms and the appearance of new 210, 180 and 170 kDa variant glycoprotein forms which were not found on parental cells. This PMA effect was not accompanied by the programmed cell death and was markedly blocked by a nonselective protein kinase (PK) inhibitor isoquinoline sulfonamide H7. Modulation of CD45 by phorbol esters might serve as an in vitro model for an additional insight into the function of CD45 in hematopoietic cells.

Key words: pre-B leukemic cells, CD45 isoforms, PMA, protein kinase inhibitor H7.
pp. 205-211

Download full text in PDF format

Circulation of progenitor cells after intensive chemotherapy followed by combination G-CSF and EPO in breast carcinoma

S. Filip, J. Vaòásek, M. Bláha, J. Vávrová

Department of Oncology and Radiotherapy, Charles University Hospital, 502 36 Hradec Králové, Czech Republic;
2nd Department of Internal Medicine, Charles University Hospital, Hradec Králové, Czech Republic;
Military Medical Academy JEP, Hradec Králové, Czech Republic

Hematologic effects of granulocyte colony-stimulating factor (G-CSF) and erythropoietin (EPO) combination after priming intensive chemotherapy in the treatment of female breast carcinoma are presented. In a previous group treated with G-CSF alone, 36% of patients became anemic and had to be transfused for correction of their anemia.
To the present study 11 consecutive patients with different stages of breast carcinoma were admitted. All were given priming intensive chemotherapy (epirubicin 150 mg/m2 and cyclophosphamide 1300 mg/m2) followed by subcutaneous application of G-CSF at a dose of 5 µg/kg/day and EPO 250 IU/kg/day. In cases where leucocyte counts dropped below 1 x 109/l and hemoglobin levels fell to 85 g/l administration of growth factors was started. The therapy was stopped when normal leucocyte count reached 4 x 109/l for G-CSF and hemoglobin level rose to 115 g/l for EPO.
Our results show significant difference between MNC/Þl (min.), CD34+ cells/µl (min.), CFU-GM/ml (min.), BFU-E/ml (min.) and MNC/µl (max.), CD34+cells/µl (max.), CFU-GM/ml (max.), BFU-E/ml (max.) p < 0.01, with mean peak values of 16.9-fold for circulating MNC/µl, 7.8-fold for CD34+ cells/µl, 23.4-fold for CFU-GM/ml and 28.7-fold increase for BFU-E/ml. Side effects were minimal, no infectious complications occurred, body temperature did not rise over 38°C and no corrections of anemia were needed.
It is concluded that the administration of G-CSF plus EPO combination following intensive chemotherapy reduces hematologic toxicity and induces large amount of hemopoietic progenitors suitable for autologous transplantation in women with breast carcinoma.

Key words: G-CSF and EPO, progenitor cells, CD34+ cells, intensive chemotherapy, breast carcinoma.
pp. 212-218

Download full text in PDF format

Cathepsin B, thiols and cysteine protease inhibitors in squamous-cell lung cancer

E. Køepela, J. Procházka, B. Kárová, J. Èermák, H. Roubková

Department of Molecular and Cellular Pneumology, Clinic of Pneumology and Chest Surgery, Medical Faculty Hospital Bulovka, 180 71 Prague 8, Czech Republic;
Department of Chest Surgery, Clinic of Pneumology and Chest Surgery, Medical Faculty Hospital Bulovka, Prague, Czech Republic;
Department of Pathology, Medical Faculty Hospital Bulovka, Prague, Czech Republic

We investigated activities of the cysteine protease cathepsin B (CB; EC, the levels of reduced glutathione (GSH) and cysteine and the activity of gamma-glutamyltransferase (gamma-GT; EC in squamous-cell lung carcinoma (SQCLC) and the lung parenchyma specimens from surgically treated patients. The basal CB activity, assayed in tissue extracts in the absence of exogenous activators, was significantly higher in SQCLC compared to the lung. The residual CB activity, remaining in tissue extracts after preincubation at 37°C, was not any longer significantly different in SQCLC and the lungs. The inhibited CB activity, calculated as the difference between the basal and residual CB activities, was significantly higher in SQCLC compared to the lung. In the case of the cysteine protease cathepsin C (CC; EC, neither the basal nor the residual nor the inhibited CC activities in SQCLC and the lung were significantly different. Compared to CC, the powerfulness of endogenous cysteine protease inhibitors to inhibit CB was much higher in both SQCLC and the lung. The cysteine protease inhibitors from SQCLC and the lung which effectively inhibited CB could be related to the inhibitors with an apparent Mr ranging from 10 000 to 30 000. Isoelectric focusing studies indicated significant differences in the progress of inhibition of the activity of CB isoforms in SQCLC and lung parenchyma extracts. The levels of both GSH and Cys were significantly higher in SQCLC compared to the lung and the level of GSH was significantly higher in Stage III tumors compared to Stage I tumors. The activity of gamma-GT was not significantly different in SQCLC and the lung but it was significantly higher in Stage I tumors compared to Stage III tumors and showed a significant negative correlation with GSH level in SQCLC. Dithiothreitol did not increase the basal activity of CB from SQCLC and the lung which indicates that reversibly oxidized forms of CB do not accumulate in the tumors and the lungs. The basal activity of CB from SQCLC and the lung was competitively inhibited by Cys. Moreover, increasing Cys concentrations had a modulatory effect on the basal activity of CB from SQCLC and the lung which was featured by Cys-induced inhibition of CB activity and by subsequent Cys-effected recovery of CB activity from its previous inhibition by Cys.

Key words: Squamous-cell lung carcinoma, cathepsin B, cathepsin C, gamma-glutamyltransferase, glutathione, cysteine, cysteine protease inhibitors.
pp. 219-239

Download full text in PDF format

Antiproliferation activity and the mechanism of action of 9-bromo-5-morpholino-tetrazolo[1,5-c]quinazoline - potentional anticancer drug

S. Jantová, ¼. Ružeková, Š. Stankovský, K. Špirková

Department of Biochemistry and Microbiology, Faculty of Chemical Technology, Slovak University of Technology, 812 37 Bratislava, Slovakia;
Department of Mutagenesis and Chemical Carcinogenesis, Cancer Research Institute of the Slovak Academy of Sciences, Bratislava, Slovakia;
Department of Organic Chemistry, Faculty of Chemical Technology, Slovak University of Technology, Bratislava, Slovakia

9-Bromo-5-morpholino-tetrazolo[1,5-c]quinazoline (BMTQ) at the two highest tested concentrations (74.6; 29.8 µmol//l) induced retarded cytotoxic effect. After 24 hours of culturing 23.1-98.8% of the cell population proliferated but after 48 and 72 hours 6.4-80.4% of the cell population degenerated. Other concentrations induced toxicity that was concentration- and time-dependent. The cytolytic concentrations of BMTQ induced integrity damage of cytoplasmatic membrane. The inhibition of cell cycle and the elevated content of proteins in the cell exposed to the cytotoxic concentrations of BMTQ suggest that the cells synthesize protein without entering into mitosis and that dying cells are in the S-phase before death. BMTQ induced 1.75-3.01-times increase of the level of ssDNA in comparison with the control.

Key words: Cytotoxicity, quinazoline derivatives, cell cycle, unbalanced growth, DNA single-strand breaks.
pp. 240-246

Download full text in PDF format

DNA-synthesis inhibition and repair DNA-synthesis in CHO Ade- C cells: An alternative approach to genotoxicity testing

D. Slameòová, A. Collins, L. Wsólová, E. Papšová, A. Gábelová, M. Dušinská

Cancer Research Institute of Slovak Academy of Sciences, 812 32 Bratislava, Slovakia;
Rowett Research Institute, Aberdeen, U.K.;
Institute of Preventive and Clinical Medicine, Bratislava, Slovakia

We describe an alternative assay to determine genotoxicity. Its main feature is that it combines two measures in a single experiment; the inhibition of replicative DNA synthesis together with the stimulation of DNA repair. We show that, in tests of four different genotoxic agents, the assay gives results that are entirely consistent with what is known about the mode of action of these agents. In addition, we have demonstrated that chemical carcinogens requiring metabolic activation can be examined using a standard procedure of incubation with a microsomal activating fraction. We consider the combined assay for DNA synthesis inhibition and repair synthesis to be a useful way for the rapid prescreening of chemicals suspected of genotoxic activity on the level of mammalian cells.

Key words: CHO, Ade-C cells, UV-light, 4-nitroquinoline N-oxide, N-methyl-N-nitrosourea, benzo[a]pyrene.
pp. 247-252

Download full text in PDF format

The influence of melatonin on metabolic changes in female rats induced by continuous irradiation and/or administration of 7,12-dimethylbenz/a/anthracene

I. Ahlers, P. Solár, E. Ahlersová, M. Kassayová, B. Šmajda

Institute of Animal Physiology, Faculty of Science, P.J. Šafárik University, 041 67 Košice, Slovakia

Metabolic profile is an important biological marker of neoplastic processes not only in the tumor itself but also in the host organism. The neurohormone melatonin has been implicated in experiments as an oncostatic agent.
Female Wistar:Han SPF rats (Velaz, Prague, Czech Republic) were irradiated continuously for 15 days using a daily gamma rays dose of 96 mGy. At the end of exposure one group of rats was administered 5 mg/kg b.w. of dimethylbenz/a/anthracene (DMBA) intragastrically. During the period of exposure to ionizing radiation a part of the animals was supplied with melatonin (M) at a concentration of 20 µl/ml in drinking water. Selected parameters of lipid and carbohydrate metabolisms and levels of selected hormones were determined 2, 30 and 100 days post-irradiation. The irradiation itself caused only small changes in tissue lipids. The application of a single low dose (subthreshold from the point of view of induction of mammary tumors) of DMBA caused more pronounced changes in nonirradiated animals; of the changes observed an increase in lipids in the liver, triacylglycerols (TG) in the thymus and decrease in myocardial glycogen predominated. The intake (by drinking) of exogenous M prevented the biochemical pattern of fatty liver in animals administered DMBA in both groups, irradiated and nonirradiated. A prolonged effect of exogenous M, demonstrated by prevention of increase in TG in the thymus and of irradiated animals caused by administration of DMBA, was observed.
The mechanism of metabolic effect of M is not known. Additional experiments are needed to explain the relationship between the beneficial effect of M on metabolic changes and its presumable oncostatic effect in rats.

Key words: Irradiated rats, dimethylbenz/a/anthracene, melatonin, metabolic changes.
pp. 253-257

Download full text in PDF format

The cytogenetic identification of high risk members in colorectal cancer families

B. Jyothish, Ravindran Ankathil, R. Chandini, B. Vinodkumar, G. Sunil Nayar, D. Dinesh Roy, Jayaprakash Madhavan, M. Krishnan Nair

Division of Cancer Research, Regional Cancer Centre, Trivandrum - 695 011 India;
Division of Radiotherapy, Regional Cancer Centre,Trivandrum, India

Family history of colorectal cancer is recognized as a risk factor for the disease and the development of colorectal cancer represents a suitable model for illustrating multistep tumor development. Bleomycin induced chromosome sensitivity studies were done in 7 colorectal cancer families consisting of 12 colorectal cancer patients and their 34 first degree relatives and 12 sporadic colorectal patients for comparison and identification of high risk family members with genetic instability. All patients and 4 unaffected relatives showed increased bleomycin sensitivity, which might be due to defective DNA repair system. These four relatives may be classified as high risk (without cancer at present) individuals. The study is being continued in more number of familial colorectal cancer patients and their relatives to arrive at definite conclusions.

Key words: Bleomycin, familial colorectal cancer, genetic susceptibility, risk factors.
pp. 258-262

Download full text in PDF format

ABVD chemotherapy of Hodgkin's disease

Z. Molnár, T. Schneider, E. Várady, T. Fleischmann

National Institute of Oncology, 1122 Budapest, Hungary

The authors report on their results in ABVD therapy, which was given by 91 patients with Hodgkin's disease as first-line treatment. 78 patients (85%) achieved complete, 10 (11%) partial remission, 3 (4%) did not respond to therapy. In the follow-up period (36-223 months) 6 patients (7%) died because of the progression of Hodgkin's disease. Serious side effects or treatment-related death did not occur. Based on their results of ABVD, the authors find ABVD chemotherapy effective as first-line treatment of Hodgkin's disease.

Key words: Hodgkin's disease, chemotherapy ABVD.
pp. 263-265

Download full text in PDF format

Cellular dysplasia in acquired cystic renal disease: Comparison of histomorphometrically gauged nuclear parameters in normal kidneys, renal cell carcinomas and acquired cystic kidneys

E. Sabo, I. Misselevich, J. Behar, O. Nativ, J.H. Boss

Department of Pathology, Bnai-Zion Medical Center and The Bruce Rappaport Faculty of Medicine, Technion - Israel, Institute of Technology, 31048 Haifa, Israel;
Department of Urology, Bnai-Zion Medical Center and The Bruce Rappaport Faculty of Medicine, Technion - Israel, Institute of Technology, Haifa, Israel

Nuclear parameters were assessed by computer-assisted image analysis in the the cells of abnormal epithelial formations in the acquired cystic kidneys of two dialysis patients, the proximal and distal tubules of a normal kidney and two well differentiated renal cell carcinomas. One acquired cystic kidney contained many small clear celled foci and an 0.9 cm-size clear celled lesion and the second one a papillary microadenoma. The clear celled lesion was cytologically indistinguishable from the carcinomas. The histomorphometrically gauged nuclear parameters were maximal and minimal ferret diameters, averaged ferret diameter, aspect ratio, shape factor, area, volume and specific length and width. Statistical evaluation evidenced that the nuclear area, volume, aspect ratio and shape factor allowed for the distinction between benign and malignant epithelial structures. The medians of the nuclear parameters of atrophic tubules, cysts, clear celled foci, papillary adenoma and clear celled lesion in the two acquired cystic kidneys deviation from those of normal renal tubules and, in increasing order of disparity, approached those of the carcinomas.

Key words: Acquired cystic kidney, renal cell carcinoma, carcinogenesis, histomorphometry.
pp. 266-271

Download full text in PDF format

Reduction in the cumulative incidence rate of cervical cancer by one life time selective screening

A. Juneja, N.S. Murthy, R.K. Tuteja, S. Sardana, D.K. Das

Institute of Cytology and Preventive Oncology, Maulana Azad Medical College, B.S.Z. Marg, New Delhi, 110002 India;
Department of Statistics, M.D. University, Rohtak, India

Mass scale cervical cytology which is the most accepted strategy for the control of cervical cancer cannot be undertaken in developing countries in view of paucity of resources, hence a need arises to examine alternate strategy. The present exercise attempts to study the reduction in cumulative incidence rate of cervical cancer by one life time selective screening. The results revealed that cumulative incidence rate (CIR) of cervical cancer per 100 000 in cohort of women during the age of 20 to 64 years was found to be 2555.0 in the absence of screening. One life time selective screening at the age of 40 and 45 years showed the reduction of 11.6 and 17.2% in CIR respectively where as respective estimates in case of complete screening at mentioned age groups were found to be 21.5% and 25%. In order to further conserve the resources the strategy seems to be optimum for developing countries.

Key words: Cancer cervix, selective screening.
pp. 272-274

Download full text in PDF format

Report on the 4th Central European Lung Cancer Conference, September 26-29, 1996, Gdañsk, Poland

pp. 275-276

Download full text in PDF format