Electronic Library of Scientific Literature


Volume 47 / No. 3 / 2000



Trends in the incidence of non-melanoma skin cancer in Slovakia, 1978–1995

I. Pleško, G. Severi, A. Obšitníková, P. Boyle

National Cancer Registry, National Cancer Institute, 833 10 Bratislava, Slovak Republic, e.mail: plesko@computel.sk;
Department of Epidemiology, Cancer Research Institute of the Slovak Academy of Sciences, Bratislava, Slovak Republic;
Division of Epidemiology and Biostatistics, European Institute of Oncology, Milano, Italy

Non-melanoma skin cancer (NMSC) incidence in Slovakia in the period 1978–1995 was analyzed. A total of 38 629 microscopically confirmed NMSC cases (19 600 in males and 19 029 in females) were registered by the National Cancer Registry: 31 714 (82.1%) were basal cell carcinomas (BCC), 6396 (16.6%) squamous cell carcinomas (SCC) and only 519 (1.3%) other NMSC. Age–standardized rates of NMSC increased in the given period by 59.1% in males and 58.5% in females. The greatest increase was observed for BCC, 70.4% and 65.0% in males and females respectively, the smaller for SCC, 13.5 and 18.8%. In the period 1993–1995 age-standardized incidence rates per 100 000 were 38.0 for BCC, 6.7 for SCC and 45.5 for all NMSC in males and 29.2, 3.8 and 33.6, respectively in females. The observed marked increase of incidence with age was particularly pronounced for SCC. In both sexes, head and neck was the most common localization of BCC and SCC (84.2 to 74.7%), followed by trunk for BCC (17.0% in males and 10.8% in females) and by upper limbs for SCC (with 11.6% in males and 12.5% in females). Very fast increase of BCC incidence over time, its slower increase with age as compared to SCC incidence and body-site distribution suggest that BCC etiology is much more similar to melanoma etiology than SCC one. Registration of NMSC in relation to changes in possible risk factors (i.e. sun exposure/protection, ozone layer decrease) is important to study the mechanism of disease occurrence and to support public health interventions.

Key words: Skin cancers, non-melanoma, epidemiology, incidence, time trends.
Neoplasma, 47, 3, 2000, 137-142


Expression of Bcl-2 in dysplastic and neoplastic cervical lesions in relation to cell proliferation and HPV infection

S. Brychtová, T. Brychta, L. Kotršová, R. Pilka, M. Tichý, V. Tichá, Z. Kolář

Institute of Pathology, Faculty of Medicine, Palacký University, 77515 Olomouc, Czech Republic, e-mail: brychtov@ tunw.upol.cz;
Laboratory of Molecular Pathology, Faculty of Medicine, Palacký University, Olomouc, Czech Republic;
Faculty of Sports, Palacký University, Olomouc, Czech Republic;
Institute of Gynecology, University Hospital, Olomouc, Czech Republic

Expression of the bcl-2 gene has been shown to effectively confer resistance to programmed cell death in a variety of tumors. The bcl-2 proto-oncogene is involved in the development of human follicular lymphomas and also in a number of solid tumors such as carcinomas of prostate, breast, lung and GIT. The present study was designed to analyze the role of Bcl-2 expression in cervical intraepithelial squamous neoplasias (CIN) and cervical invasive carcinomas. Special attention was given to the association of Bcl-2 expression with the grade of the lesion, proliferative activity (expression of nuclear antigen of proliferative cells – PCNA) and human papillomavirus (HPV) DNA positivity. We examined tissue samples obtained from 86 women with varying degrees of cervical disease. Bcl-2 and PCNA were investigated using immunohistochemical staining and detection of HPV DNA was performed by hybridization in situ. Increased Bcl-2 expression was observed in advanced degrees of dysplasia and in carcinomas. We found a strong association between the presence of Bcl-2 in pathological epithelium with both the degree of dysplasia and the proliferative activity. We also observed a significant correlation between the amount of Bcl-2 positive lymphocytes infiltrating the lesions and the degree of disease. We, therefore, suggest that Bcl-2 expression in these lymphocytes may influence the antiviral or antitumor immune response. On the other hand we did not detect any significant correlation between the Bcl-2 oncoprotein and the presence of HPV. These results indicate that Bcl-2 may play an important role in the development of cervical cancer.

Key words: Cervical cancer, human papillomavirus, Bcl-2, proliferation, carcinogenesis.
Neoplasma, 47, 3, 2000, 143-147


Loss of heterozygosity on chromosome 5 in vicinity of the telomere in gamma-radiation-induced thymic lymphomas in mice

M. Sitarz, E. Wirth-Dzieciolowska, P.Demant

Department of Genetics and Laboratory Animal Breeding, M.Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, PL 02-781 Warsaw, Poland, e-mail: jontek@coi.waw.pl;
Division of Molecular Genetics, The Netherlands Cancer Institute, Amsterdam, The Netherlands

Analysis of the loss of heterozygosity at the D5Mit143 locus was done for thymic lymphomas induced by gamma-irradiation of mice from two reciprocal backcrosses (BALB/c x CcS-13)F1 x BALB/c and (BALB/c x CcS-13)F1 x CcS-13. BALB/c mice are susceptible to gamma-ray induction of lymphomas. The CcS-13 strain is one of 20 CcS/Dem (CcS) series of recombinant congenic strains, and the CcS-13 mice are resistant to gamma-radiation-induced lymphomas [1, 8].
Our preliminary tests show 50% (6/12) frequency of allelic loss at the D5Mit143 locus in thymic lymphomas induced by gamma-irradiation of the mice from (BALB/c x CcS-13)F1 x BALB/c backcross. Yet, in gamma-radiation-induced lymphomas from the backcross made in opposite direction, namely, (BALB/c x CcS-13)F1 x CcS-13, the analysis with the D5Mit143 marker revealed low incidence of the loss of heterozygosity, 6.7% (1/15). The D5Mit143 locus resides in the distal part of chromosome 5, close to the telomere.
Allelic loss of heterozygosity at the D5Mit143 locus showed strain specificity. In each case, the lost allele derived from the CcS-13 resistant strain.
Our current results and previously done linkage analysis [8] let us to suspect existence of a putative tumor suppressor gene for gamma-radiation-induced lymphoma at the region of murine chromosome 5.

Key words: Loss of heterozygosity (LOH), gamma-radiation-induced thymic lymphomas, recombinant congenic strains (CcS/Dem).
Neoplasma, 47, 3, 2000, 148-150


The expression of apoptosis-related proteins and the apoptotic rate in glial tumors of the brain

J. Ehrmann Jr., P. Řiháková, A. Hlobilková, M. Kala, Z. Kolář

Institute of Pathology, Laboratory of Molecular Pathology, Center of Molecular Biology and Medicine, Faculty of Medicine, Palacký University, 775 15, Olomouc, Czech Republic, e-mail: erman@tunw.upol.cz;
Department of Neurosurgery, Faculty of Medicine, Palacký University, Olomouc, Czech Republic

Modern molecular biology methods allow a more precise analysis of the biological characteristics of tumors and, consequently, a more precise treatment plan. The determination of apoptotic rate and expression of apoptosis-related proteins belong among the important prognostic/diagnostic markers in many tumors. The validity of these factors had not yet been sufficiently analyzed in astroglial tumors. The aim of this work was therefore to study mutual relationships between apoptotic rate, expression of apoptosis-regulating proteins and some clinical and histopathological data.
The TUNEL method was used for the determination of apoptosis in 44 astroglial tumor specimens.The percentage of TUNEL positive cells was expressed by the TUNEL index (TI). The TI data was compared with the immunohistochemically detected expression of proteins involved in apoptosis (BCL-2, FAS, FAS-L, and caspase 1), with grading, age, proliferative activity (assessed by PCNA expression analysis) and overall survival of patients. The statistical evaluation of results was done by two-way sample analysis of variance.
We have demonstrated significantly higher values of both TI and expression of FAS-L and caspase 1 in low grade tumors, which were characterized by a longer survival, lower average age and a lower expression of PCNA. FAS-L expression correlated significantly with the expression of the caspase 1. No significant difference was found between the expression of BCL-2 and FAS. These results suggest that the determination of TI in astroglial tumors may be an important prognostic marker. The expression of FAS-L and caspase 1 in low grade astroglial tumors could indicate the increased readiness to apoptosis via the FAS/FAS-L cascade.

Key words: Glial tumors, apoptosis, prognosis.
Neoplasma, 47, 3, 2000, 151-155


Detection of Epstein-Barr virus in Hodgkin’s lymphoma (patients in the Czech Republic)

J. Mačák, B. Habanec, P. Fabián

Institute of Pathology of Medical Faculty, Palacký University, 775 15 Olomouc, Czech Republic, e-mail: MACAK@risc.upol.cz;
The 2nd Institute of Pathology of Medical Faculty, Masaryk University, Brno, Czech Republic

The frequency of EBV demonstrated in patients with Hodgkin’s lymphoma (HL) shows geographical variability. In the present study, we investigated the frequency of EBV in HL patients in the Czech Republic. The presence of EBV was determined by immunohistochemistry (IHC) with anti LMP-1 antibody and by in situ hybridization (ISH) method for EBERs. We studied 142 cases with HL. The age of patients ranged from 4 to 82 years. The male to female ratio was 1.2 (males 55.6%). In the series of 142 patients 47 (33%) positive cases were found. The incidence of EBV-positive results was significantly, higher in males than in females (70.2 vs. 29.7%, = 0.023). Five patients were found in the age group below 10 years. They were positive with LMP-1 antibody and for EBERs in ISH method. The same results were discovered in two patients above the age of eighty. The most frequent histologic types of HL were nodular sclerosis (64 cases) and mixed cellularity (62 cases), respectively. The former type contained 16 EBV-positive cases (25%) and the latter 24 (38%) positive cases. The lymphocyte depletion type 2 (67%); lymphocyte rich type 5 (38%). EBV-positivity examined by ISH and IHC methods determined not only diagnostic Hondgkin cells and Reed-Sternberg cells but also small lymphocytes. In IHC method were small lymphocytes positive in 11 cases, more sensitive ISH revealed 32 positive cases.

Key words: Hodgkin’s lymphoma, EBV, immunohistochemistry, in situ hybridization.
Neoplasma, 47, 3, 2000, 156-161


Application of cytogenetic endpoints and Comet assay on human lymphocytes treated with vincristine in vitro

N. Kopjar, V. Garaj-Vrhovac

Institute for Medical Research and Occupational Health, HR-10 000 Zagreb, Croatia, e-mail: nkopjar@imi.hr

The genotoxic potential of vincristine is assessed on human peripheral blood lymphocytes following administration of the drug at a dose 0.0875 µg/ml by use of single cell gel electrophoresis – Comet assay (SCGE), analysis of structural chromosome aberrations (CA), micronucleus assay (MN) and sister chromatid exchange (SCE) analysis. In vitro treatment of human lymphocytes with vincristine was performed on cells in G0 phase, as well on lymphocyte cultures 24 hours after stimulation with mitogen phytohemagglutinine. For the Comet assay at 24, 48 and 72 h the treated cells were embedded in agarose on slides, lysed with alkaline lysis solution and exposed to an electric field. DNA migrated within the agarose and formed comets whose length depends on the amount of DNA damage. For the analysis of structural CA cells were grown on F-10 medium for 48 hours, and for MN and SCE analysis for 72 hours. The results on SCGE showed an increase in tail length compared to control both in cells treated in G0 and in cells treated 24 h after mitogen stimulation. The amount of DNA damage was higher in cells treated with vincristine 24 h after mitogen stimulation. Administered concentration of drug caused total inhibition of lymphocytes growth in 72-h cultures for MN and SCE analysis indicating strong microtubule distruptive effects of vincristine. Analysis of structural CA reveals chromatid breaks and acentric fragments as the main aberration types both in cells treated in G0 and in cells treated 24 h after mitogen stimulation. Number of these aberrations was higher in cells treated in G0 phase. Results obtained in this study by use of different cytogenetic endpoints confirmed that vincristine exhibits both aneugenic and clastogenic effects on human lymphocytes.

Key words: Human lymphocytes, vincristine, in vitro, comet assay, cytogenetic endpoints.
Neoplasma, 47, 3, 2000, 162-167


Dexamethasone does not enhance antileukemic activity of cladribine in mice with leukemias L1210 and P388

T. Robak, A. Szmigielska

Department of Hematology, Medical University of Łódź, 93-513 Łódź, Poland, e-mail: robaktad@ psk2.am.lodz.pl

Combination of corticosteroids with new purine analogs such as cladribine 2-chlorodeoxyadenosine, (2-CdA) and fludarabine (FAMP) is controversial. The possibility of potentiation of antineoplastic activity of 2-CdA or FAMP by corticosteroids has not been documented so far. On the other hand, such combination may increase immunosuppression and the risk of infections. The aim of our study was to evaluate the influence of 2-CdA and dexamethasone (DEX) on the survival time of mice bearing lymphoid leukemias L1210 and P388.
CD2F1 strain mice (132 male) were used in the experiment. The animals were injected i.p. on day 0 with 106 leukemic cells. The drugs were given on days 1–5 i.p. in the following concentrations: 2-CdA – 20 mg/kg, DEX 1.25 mg/kg, DEX 2.5 mg/kg, DEX 5 mg/kg, DEX 10 mg/kg, alone and in combination. The animals were observed daily for survival for a minimum of 30 days. The efficacy of the therapy against leukemia (defined as increase in lifespan – ILS) was assesed as the percentage of the median survival time (MST) of the treated group(t) to that of the control group(c): ILS(%) = (MSTt/MSTc)100.
The survival time of mice bearing L1210 or P388 leukemia treated with both drugs simultaneously was not longer than that of mice treated with either of drugs alone. Combination of 2-CdA and DEX in doses 5 and 10 mg/kg resulted in decrease of survival time of animals bearing P388 leukemia as compared with the control group without any treatment.
Our study revealed that combination of 2-CdA with DEX in both leukemias is not more effective than 2-CdA alone. These results may indicate that routine addition of corticosteroids to purine analogs in the treatment of lymphoid malignancies is not warranted.

Key words: 2-chlorodeoxyadenosine, cladribine, dexamethasone, corticosteroids, murine leukemias, interactions.
Neoplasma, 47, 3, 2000, 168-171


Comparison of three in vitro assays at evaluation of IC50 of acetylsalicylic acid, ferrous sulfate, amitriptyline, methanol, isopropanol and ethylene glycol in human cancer cells HeLa

K. Ruppová, L. Wsólová, M. Urbančíková, D. Slameňová

Institute of Preventive and Clinical Medicine, 833 01 Bratislava, Slovak Republic;
Cancer Research Institute, Slovak Academy of Sciences, Bratislava, Slovak Republic, e-mail: exonslam@savba.sk

Evaluation of the 50% inhibitory concentration (IC50) of acetylsalicylic acid, ferrous sulfate, amitriptyline, methanol, isopropanol and ethylene glycol was done on human cancer cells cultured in in vitro conditions. Three different in vitro assays were used in this study: the plating efficiency test, the microprotein test and the neutral red uptake test. Obtained results were evaluated by statistical methods. All used methods seem to be useful for screening a cytotoxic potential of the tested chemicals. The knowledge of cytotoxic effects of frequently used chemicals on mammalian cells is important not only for necessary in vitro genotoxicity and carcinogenicity studies but also for assessing the toxicity of chemicals to find out possible hazards to the human health. Results presented in this paper underline the usefulness of the wider methodological approach for the comparison of the different endpoints as well as a necessity for selection of a battery of in vitro cytotoxicity tests allowing to estimate the possible harmful effects of xenobiotics.

Key words: HeLa cell line, plating efficiency, microprotein method, neutral red assay, in vitro toxicity, IC50 values.
Neoplasma, 47, 3, 2000, 172-176


Adjuvant clodronate therapy in patients with locally advanced breast cancer – long term results of a double blind randomized trial

J. Mardiak, L.Bohunický, J. Chovanec, T. Šálek, I. Koza, the Slovak Clodronate Collaborative Group

National Cancer Institute, 833 10 Bratislava, Slovak Republic, nci@bts.sk

Between March 1990 and May 1993 seventy three patients with previously untreated breast cancer, Stage III or IV without osseal metastases were randomized to sodium clodronate 1600 mg daily p.o.(arm A = 37 patients) or placebo (arm B = 36 patients) over 2 years, additionally to standard therapy. Ten patients were not evaluable for response because of short duration of therapy (less than 2 months). Bone metastases developed in 30% of patients in arm A and 23% patients in arm B. Median time to appearance of bone metastases was 13 months in arm A and 28 months in arm B. Non-bone metastases appeared in 48% patients in arm A and in 48% patients in arm B. Time to development of non-bone metastases was 20 months in arm A and 16 months in arm B. Five-year survival was 41% in arm A and 39% in arm B. There were no significant differences between the treated and control arms.

Key words: Biphosphonates, clodronate, untreated breast cancer.
Neoplasma, 47, 3, 2000, 177-180


The impact of radiotherapy on the incidence and time of occurrence of local recurrence in early-stage breast cancer after breast conserving therapy

J. Fodor, T. Major, C. Polgár, J. Tóth, G. Németh

Department of Radiation Therapy, National Institute of Oncology, Budapest, H-1122, Hungary, e-mail: fodor@oncol.hu;
Department of Human and Experimental Tumor Pathology, National Institute of Oncology, Budapest, Hungary

There is still little information on the delay of local recurrence after conservatively treated and irradiation breast cancer. To evaluate the impact of radiation therapy (RT) on the incidence and on the time of occurrence of ipsilateral breast tumor recurrence (IBTR), we reviewed the treatment results in 415 women with UICC Stage I or II unilateral breast cancer. All underwent breast conserving surgery (BCS) and full axillary dissection between 1983 and l987. Out of them 309 patients were irradiated and 106 were not. The median dose of RT was 50 Gy in five weeks to the whole breast. Systemic therapy, when it was given, consisted of 6-cycles of CMF for node positive premenopausal women and 20 mg tamoxifen for three years for postmenopausal women. The median follow up time was 120 months in survivors. The 10-year actuarial IBTR rate was 36.6% for the nonirradiated and 9.1% for the irradiated women (p = 0.0000); 48.6% for patients treated with CMF and 4.2% for those treated with CMF plus RT (p = 0.0051); 29.0% for patients treated with tamoxifen and 7.9% for those treated with tamoxifen plus RT (p = 0.0318). The patient’s age and the presence of an extensive intraductal component (EIC) were both highly associated with the likelihood of tumor recurrence in the treated breast. Patients under 41 years of age had an actuarial 10-year IBTR rate of 75% without RT and 17.1% with RT (p = 0.0006). Women with an EIC positive tumor had an IBTR rate of 88.9% when RT was not given and 27.2% when RT was given (p = 0.0003). In invasive lobular cancer, irradiated patients had a IBTR rate of 2.3%, compared to 53.2% for nonirradiated patients (p = 0.0008). RT resulted in a significant delay in the appearance of IBTR (p = 0.0250) and the median time was increased by 20.0 months.
We conclude that RT has the property of not only preventing but also delaying IBTR. In invasive lobular tumors the risk of IBTR is very high when RT is omitted, but BCS plus radiation therapy is effective treatment. Patients wih EIC positive tumor are at high risk of IBTR even when a median dose of 50 Gy is given to the whole breast.

Key words: Breast cancer, conservative treatment, irradiation.
Neoplasma, 47, 3, 2000, 181-186


Influence of previous treatment on palliative effect of HDR brachytherapy in advanced esophageal cancer

B. Białas, T. Rutkowski, M. Fijałkowski, A. Rembielak

Brachytherapy Department, Centre of Oncology Maria Skłodowska-Curie, Gliwice, Poland, e-mail: tomr@instonko.gliwice.pl

General effectiveness and influence of previous treatment on value of palliative HDR brachytherapy were assessed in 35 patients with advanced esophageal cancer treated from 1992 till 1997 with brachytherapy HDR (BT). Twelve of them were treated only with BT, 11 received previously chemoradiotherapy (CHTT), 12 teleradiotherapy (TT). BT appeared to be effective method of palliation. No significant differences in effectiveness of BT in analyzed groups were observed. Sever complications were observed in 9 cases (26%), and that in patients treated previously. Brachytherapy seems to be efficient after previous treatment, however, in this case, the risk of complications increases.

Key words: HDR brachytherapy, esophageal cancer, palliative treatment.
Neoplasma, 47, 3, 2000, 187-190


Male breast cancer. Does the prognosis differ compared to female?

K. Herman, W. Łobaziewicz, P. Skotnicki, J. Fortuna, T. Kusy, T. Leśniak

Department of Surgical Oncology, Cancer Center, 31-115 Kraków, Poland, e-mail: hermank@polbox.com;
County Hospital, Sucha Beskidzka, Poland;
Resovian Cancer Center, Poland;
Regional Cancer Center, Bielsko-Biała, Poland

Due to the low incidence of breast cancer in males there are not many reports in the literature. In this study we analyzed results of treatment in 65 breast cancer males, who had been treated in one institution. Radical surgery was performed in 45 patients. Lymph node metastases were found in 25 patients (55.5%), the tumor was usually moderately differentiated (21 pts – 46.7%). Median survival after radical surgery was 73 months compared to 38 months for nonsurgical patients (p < 0.0001). In the group of males after radical surgery the results of 5-, 10- and 15-year survival rates were 69.8, 59.7 and 31.3% respectively. Comparable analysis of two subgroups of patients with favorable (T1 or T2, N0, grade I or II) and unfavorable (T3 or N+ or grade III) prognostic factors was also performed. In the first subgroup the 5-, 10- and 15-year survival rates were 90, 77.4 and 62%, compared to 61.8, 23.1 and 23.1% for the second subgroup. The multivariate analysis showed grading and node status as the strongest parameters influencing survival. Relative risk of death was over 3 times higher for nodal metastases and near 3 times higher for high grade carcinomas (p < 0.01), compared to patients without metastases and low grade of tumor. Similar analysis was performed when 45 males were compared to 500 selected women, with similar clinical parameters (age, node status, grading). Again, data indicated grading and lymph node status as the strongest prognostic factors. It was not unlikely, that gender had some influence on prognosis, when relative risk of death for males was over 1.5 times higher than for females, but this result was not clearly significant (p < 0.1). The question, whether male breast cancer prognosis is worse then in female remains open. Multiinstitutional prospective studies are needed in this area.

Key words: Male breast cancer, prognosis.
Neoplasma, 47, 3, 2000, 191-195