Electronic Library of Scientific Literature
Electronic Library of Scientific Literature
Volume 44 / No. 6 / 1997
V. Leksa, Č. Altaner
Cancer Research Institute, Slovak Academy of Sciences, 833 91 Bratislava, Slovakia
Differential display technique was applied to study expression of RNA in tumorigenic and nontumorigenic cell variants of avian sarcoma virus transformed hamster cells. Methodical conditions were worked out, which allowed identifying a cDNA fragment of an unknown gene expressed in nontumorigenic cell variant only. Its role in tumor suppression remains to be determined.
Key words: Tumorigenic and nontumorigenic cells, differential display,
RNA expression.
pp. 337-341
J. Hlavatý, K. Hlubinová, V. Altanerová, J. Líška, Č. Altaner
Cancer Research Institute, Slovak Academy of Sciences, 833 91 Bratislava, Slovakia
The retrovirus vector containing Herpes simplex virus type 1 thymidine kinase (HSVtk) gene was constructed. The vector was transfected into the packaging cell line PG13. It was shown that individual transfected cells differ in the production of recombinant retrovirus and in their susceptibility to be killed by ganciclovir. Recombinant retrovirus with a gibbon envelope was able to transduce the HSVtk gene into rat glioma cells. In vivo studies confirmed the ability of intraperitoneal ganciclovir administration to influence subcutaneous and intracerebral tumors developed after injection of C6 rat glioma cells with subsequent injection of HSVtk retrovirus producing cells.
Key words: Gene therapy, brain tumor, herpes simplex thymidine kinase,
ganciclovir.
pp. 342-347
O. Babušíková, M. Glasová, J. Stašáková, J. Kusenda, E. Koníková
Cancer Research Institute, Slovak Academy of Sciences, 833 91 Bratislava, Slovakia
In our study we used for definition of leukemia/lymphoma cells a new
parameter which allows the enumeration of mean fluorescence intensity expressed
by the number of antigen molecules per cell. Quantitative immunofluorescence
using calibration microbeads was performed in 36 patients with different
acute and chronic lymphoid and myeloid leukemia and in 19 healthy volunteers.
We showed that quantitative immunophenotyping allowed the definition of
aberrant marker densities on neoplastic cells. We demonstrated under- and
overexpression of CD8 marker in CD3/CD4/CD8 complex in T acute lymphatic
leukemia and T non-Hodgkin's lymphoma and T leukemia of large granular
lymphocytes as compared to normal counterparts. We pointed out that certain
antigens (e. g. CD10, CD4, CD24) were expressed at different levels on
different cell subsets (CD10 in early B-acute lymphatic leukemia and coexpressed
in T-acute lymphatic leukemia, CD4 on T cells and monocytes, CD24 on B
cells and granulocytes in chronic myeloid leukemia). We showed that quantitative
immune fluorescence could provide new data contributing to a more precise
definition of cell differentiation. We documented the significant difference
between antigen density of early and late markers in B-cell and myeloid
malignancies. Further, we demonstrated that quantitative immune phenotyping
could help in determination of exact definition of pathologic clone in
morphologically immature leukemia population and showed that parameters
of this method are also convenient for cytoplasmic marker evaluation. In
our study we were able to demonstrate that CD45 quantitative expression
appeared to be a more informative parameter than its percentage of antigen-positive
cells as a measure of antigen expression only and we pointed out that low
and high CD45 densities enabled to differentiate between pathological clone
and residual healthy population in examined sample.
We showed that quantitative immune phenotyping could be another important
parameter for definition of leukemia phenotype suitable for detection of
minimal residual disease.
Key words: Leukemia cells, quantitative flow cytometry, immunophenotyping,
CD markers.
pp. 348-355
M.Klobušická, O.Babušíková
Cancer Research Institute, Slovak Academy of Sciences, 833 91 Bratislava, Slovakia
This study reports the immunophenotypic features of a series of 62 selected acute leukemia patients with increased incidence of argyrophilic proteins (AgNORs) at the time of initial diagnosis. Peripheral blood and bone marrow cells of patients with T-ALL, B-precursor ALL and AML were studied. The method of silver staining was used to determine the number of AgNORs per cell. Cell surface markers were detected by a standard immunofluorescence assay. To demonstrate the relationship between AgNOR quantity and cell proliferation, the expression of activation and proliferation antigens CD38 and CD71 was investigated. To characterize the immunophenotype and the discrete stages of differentiation, the wide panel of antibodies against lymphoid, myeloid and non-lineage specific antigens was used. The number of AgNORs at diagnosis ranged from 3.05 to 6.70. Immunophenotypic analysis showed a variation in CD38 and CD71 expression among different leukemia subtypes. CD71 antigen was more expressed in T-ALL than in B-precursor ALL or in AML. Notable was the relationship between increased AgNOR quantity and antigens that characterize the immaturity of leukemic cells. The association with CD7, CD2, CD5 (without CD3 membrane expression) and CD34 in T blasts was evident. High positivity of CD19, CD10, CD34 and HLA-DR in relation to the increased amount of AgNORs in B-lineage ALL was observed. The vast majority of AML patients with high numbers of AgNORs simultaneously expressed CD13, CD33, CD34 and HLA-DR. One third of AML cases coexpressed T cell marker CD7. In conclusion, the presence of increased numbers of AgNORs at diagnosis might reflect the dependence on an early stage of leukemia cell differentiation.
Key words: Acute leukemia, argyrophilic proteins, immunophenotypic
markers, cell proliferation.
pp. 356-360
N.P. Konovalova, L.M. Volkova, L.V. Tatyanenko, R.A. Kotelnikova, T.N. Yakushchenko, T.V. Kagiya
Institute of Chemical Physics, Russian Academy of Sciences, 142 432
Chernogolovka, Moscow region, Russia;
Kinki Invention Center, Kyoto, Japan
A triazole group radiosensitizer AK-2123 is shown to inhibit considerably the growth of hepatic metastases induced by the intrasplenic injection of colon adenocarcinoma cells in syngenic mice. Even an extremely low dose of the drug exhibits the antimetastatic effect. It is shown that AK-2123 injected at therapeutic dose inhibits active transport of calcium ions by the (Ca2+-Mg2+)-dependent ATP-ase. The antimetastatic effect of AK-2123 is suggested to be related, at least partially, to the inhibition of the active calcium transport.
Key words: Hepatic metastases, antimetastatic effect, radiosensitizer,
active transport, calcium ions.
pp. 361-365
Ľ. Hunáková, J. Sedlák, M. Šuliková, J. Chovancová, J. Duraj, B. Chorváth
Cancer Research Institute, Slovak Academy of Sciences, 833 91 Bratislava, Slovakia
Mevalonate pathway inhibitor lovastatin inhibited proliferation of human
multidrug-resistant promyelocytic leukemia HL-60/ADR cells in vitro, with
MRP-gene coded p190 mediated drug resistance, to a markedly lesser extent
than that of the parental drug sensitive HL-60 cells and also that of the
other human multidrug resistant (MDR-1, P-glycoprotein) myeloid leukemia
cell line HL-60/VCR. The sensitivity of the examined human leukemia cell
lines to the cytostatic activity of lovastatin correlated approximately
with the potential of lovastatin to induce the characteristic cell cycle
alteration (i.e. the accumulation of lovastatin-treated cells in the G0/G1
phase of the cell cycle).
The P-glycoprotein positive HL-60/VCR cells and the parental drug sensitive
HL-60 cells were more sensitive to this cell cycle alteration than the
HL-60/ADR multidrug resistant leukemia cells with MRP drug resistance.
Lovastatin (72 hours, 20 ľmol) induced apoptosis and cell necrosis in HL-60
cells, apoptosis but not cell necrosis in HL-60/VCR cells and neither apoptosis
nor necrosis in HL-60/ADR cells.
Key words: Lovastatin, multidrug-resistance, human leukemia, cell
line, cytotoxicity, MTT test, cell cycle analysis, MDR-1, MRP, apoptosis.
pp. 366-369
A. Rejthar, R. Nenutil
Department of Histopathology, Faculty Hospital Bohunice, 639 00 Brno, Czech Republic
The expression of cytokeratins 7, 8, 14, 18, 19 and vimentin was examined in 100 cases of ductal invasive breast carcinomas. While the predominantly diffuse immunohistological positivity of simple epithelia cytokeratins 7 (in 93), 8 (in 100), 18 (in 100) and 19 (in 97) cases represents a constant feature of these tumors, cytokeratin 14 was detected in only 36 cases which were mostly of low grade and in a focal pattern. Vimentin positivity was found in 53 intermediate and high grade tumors and, again the pattern was also rarely diffuse. The ductal carcinomas can be grouped into four classes according to vimentin and cytokeratin 14 immunoreactivity. This grouping correlates well with tumor grade and with simple histological classification of ductal breast carcinoma, consisting of the low, intermediate and high malignancy categories, as proposed here. The types of ductal carcinomas can be sorted into prognostically different subgroups, according to ICD-O morphologic terminology and commonly adopted results of morphologic and prognostic studies.
Key words: Breast cancer, histopathology, cytokeratin, vimentin,
intermediate filaments, classification, prognosis.
pp. 370-373
V. Vlčková, M. Slaninová, M.A. Morais Jr., J.A.P. Henriques, I. Fridrichová, J. Brozmanová
Department of Genetics, Faculty of Natural Sciences, Comenius University,
Bratislava, Slovakia;
Departamento de Biofisica, Centro de Biotecnologia, Universidade Federal
do Rio Grande do Sul, Porto Alegre, Brazil;
Department of Molecular Genetics, Cancer Research Institute, Slovak Academy
of Sciences, 833 19 Bratislava, Slovakia
The pso4-1 mutant of S.cerevisiae is phenotypically similar to the recA mutant of E.coli; it is sensitive to DNA cross-linking agents and defective in both recombination and mutagenesis. In this paper we have measured the effect of the recA gene expression on the frequency of mitotic crossing-over and mitotic gene conversion in response to DNA damage induced by photoactivated 8-methoxypsoralen (8-MOP + UVA), ultraviolet radiation (UV) and N-methyl-N´-nitro-N-nitrosoguanidine (MNNG). The diploid pso4-1 mutant and the repair wild type strain were transformed with the multicopy plasmid carrying the recA gene placed under the control of the ADH1 promoter. The results showed that RecA is not able to restore block in induced mitotic recombination in pso4-1 cells after DNA damaging agents used. Thus RecA protein is not able to substitute Pso4 protein in homologous mitotic recombination indicating that they have probably different functions in this process.
Key words: recA gene, pso4-1 mutant, mitotic recombination.
pp. 374-379
A. Gábelová, D. Slameňová, Ľ. Ružeková, T. Farkašová, E. Horváthová
Cancer Research Institute, 833 91 Bratislava, Slovakia
Three techniques: single cell gel electrophoresis (SCGE), alkaline elution of DNA (AE), and alkaline DNA unwinding (ADU) were chosen to compare the sensitivity among these methods in detection of DNA damage and repair in human diploid VH10 cell line after short-term exposure to hydrogen peroxide. Using SCGE technique a dose-dependent increase in DNA migration was found in cells exposed to hydrogen peroxide in concentration range from 10 micromol/l to 100 micromol/l. Alkaline DNA unwinding method detected increased level of single strand breaks (ssb) in concentration range from 25 micromol/l to 100 micromol/l of H2O2, and alkaline elution of DNA estimated increased DNA elution rate from concentration 50 micromol/l of H2O2. In a time course study to evaluate the kinetics of DNA repair, both SCGE and ADU techniques showed that the repair of DNA strand breaks is very rapid; the level of ssb in treated cells has returned to near the background level within two hours. After this time damage remaining in the DNA was in the form of oxidised bases as revealed the incubation of treated cells with specific DNA repair endonuclease, formamidopyrimidine-DNA glycosylase.
Key words: Hydrogen peroxide, single cell gel electrophoresis, alkaline
elution of DNA, alkaline DNA unwinding, VH10 cells.
pp. 380-388
A. Vachálková, L. Novotný
Cancer Research Institute, Slovak Academy of Sciences, 833 19 Bratislava, Slovakia
This work is devoted to the study of polarographic reduction of three antibiotic compounds including adriamycin, chloramphenicol and erythromycin and of a synthetic antibacterial chemotherapeutic compound - 5-nitrofurantoin. The polarographic reduction was performed in the strictly anhydrous N,N-dimethylformamide with or without alpha-lipoic acid (LA) by the means of DC polarography. The values of half-wave potentials E1/2 and parameter of potential carcinogenicity were determined for the all compounds. Adriamycin was reduced during the five-step process, other compounds were reduced in two steps. The presence of LA in a polarographic solution resulted in a new polarographic one-electron wave in the range of -1.120 V to -1.790 V vs. SCE possessing a diffuse and reversible character. Its height is linearly dependent on the LA concentration in solution. The highest parameter of potential carcinogenicity tg alpha was determined for adriamycin (0.575) which belongs among compounds classified by WHO as "probably carcinogenic to humans". The lowest determined value of parameter tg alpha belonged to 5-nitrofurantoin (0.290) which has not yet been included into the IARC classification.
Key words: Adriamycin, chloramphenicol, erythromycin, 5-nitrofurantoin,
DC polarography, alpha-lipoic acid, carcinogenicity.
pp. 389-394
M. Ścieszka, A. Danch, M. Machalski, M. Dróżdż
I Clinic of Internal Diseases, Silesian Medical Academy, 40-029 Katowice,
Poland;
Department of Biochemistry and Chemistry, Silesian Medical Academy, Katowice,
Poland
Plasma selenium concentration was assessed in 44 patients with cancer of the gastrointestinal tract (19 subjects with stomach cancer and 25 with colon cancer) and 25 age-matched healthy control subjects. Selenium concentration was determined by the fluorometric method. The observed plasma selenium concentrations in gastrointestinal cancer patients (37.0 ą 11.05 ng Se/ml or 38.4 ą 12.6 ng Se/ml in stomach or colon cancer patients, respectively) were significantly lower as compared to the healthy age-matched control group (51.4 ą 14.4 ng Se/ml). The diagnosed low selenium status may be considered as a high risk for cancer development.
Key words: Plasma selenium, stomach cancer, colorectal cancer, selenium
deficiency.
pp. 395-397
pp. 399-400
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