Electronic Library of Scientific Literature


Volume 46 / No. 5 / 1999


Tumor necrosis factor-alpha: Molecular-biological aspects: Minireview

Z. Szatmáry

Institute of Preventive and Clinical Medicine, Department of Clinical Immunology, 833 01 Bratislava, Slovak Republic

Tumor necrosis factor-alpha (TNF-alpha) a proinflammatory cytokine with multiple actions was first identified for its anticancer activity. However, TNF-alpha has a beneficial function in activation of host defense, its uncontrolled production can lead to pathological consequences. At the cellular level, it is able to exert obviously opposing effects: apoptosis and activation. It modulates survival and activates genes through various intermediates, including protein kinases, protein phosphatases, reactive oxygen intermediates, phospholipases, proteases, sphingomyelinases and transcription factors. In this review, the TNF-alpha is characterized at the molecular and cellular level (TNF-alpha mediated signal transduction is discussed in the first part, regulation of its expression in the second one), as well as methods of its determination in biological materials, giving special emphasis to the molecular-biological approach. The full understanding of the molecular mechanism of TNF-alpha will provide the basis for a pharmacological approach intended to inhibit or potentiate selected biological actions of this cytokine.

Key words: Tumor necrosis factor-alpha, TNF-alpha signal transduction, TNF-alpha expression, TNF-alpha assaying, RT-PCR, pentoxifylline.
NEOPLASMA, 46, 5, 1999, 257-266

Construction and testing of gene therapy retroviral vector expressing bacterial cytosine deaminase gene

J. Hlavatý, K. Hlubinová, Č. Altaner

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

A retroviral vector containing gene for bacterial enzyme cytosine deaminase (CD) under the control of viral LTR sequences was constructed and transfected into packaging cell line GP+envAm12. High virus titer producing single cell clone (1 x 107 cfu/ml, determined on NIH 3T3 cells) was isolated and used to transfer CD gene into human mammary carcinoma cell lines in vitro. Transduced cells exhibited high sensitivity to the antifungal drug 5-fluorocytosine (5-FC), whereas parental cells did not. Cocultivation of CD-positive and CD-negative parental cells showed bystander effect, dependent on the ratio of CD-positive cells. No enhancement of 5-FC cytotoxicity by leucovorin was observed in cells expressing cytosine deaminase.

Key words: Retroviral vector, gene therapy, bacterial cytosine deaminase, breast cancer cell lines.
NEOPLASMA, 46, 5, 1999, 267-276

Aberrant markers expression in T- and B-lymphoid and myeloid leukemia cells of different differentiation stages

O. Babušíková, E. Koníková, J. Kusenda, K. Koubek

Cancer Research Institute, Slovak Academy of Sciences, 812 32 Bratislava, Slovak Republic;
Institute of Hematology and Blood Transfusion, Prague, Czech Republic

The aim of the study was to ascertain if in T acute lymphoblastic leukemia (T-ALL), B acute lymphoblastic leukemia (B-ALL) and acute myeloid leukemia (AML) of different differentiation stages the coexistence of aberrant markers correlate with the degree of leukemic blasts maturation. We evaluated the results of surface and intracellular markers in 42 T-ALL, 86 B-ALL and 71 AML cases. A large panel of monoclonal antibodies (MoAbs) against T-cell, B-cell, myeloid cell and non-lineage specific structures has been used. Patients had dual-color flow cytometric immunophenotyping performed by FACstar flow cytometer. The correct immunological diagnosis of followed new cases before any treatment has been performed and simultaneously the presence of atypical/aberrant phenotypes has been studied and correlated with leukemia cells differentiation stage. A great deal of T-ALL and AML, in opposite to B-ALL cases, revealed a high proportion of atypical phenotypes (55, 75 and 36%, respectively), which are absent in nonleukemic cells. We found out that these atypical phenotypes were present in T-ALL, AML (not clearly in B-ALL) through all differentiation stages and so we obtained an evidence that they might represent an abnormal/atypical rather than an immature phenotype, as it was postulated till now by several authors.

Key words: Acute T-cell, B-cell lymphoblastic and myeloid leukemia, atypical/aberrant imunonophenotypes, degree of leukemia cell maturation.
NEOPLASMA, 46, 5, 1999, 277-282

Angiogenesis inhibitor TNP-470: Cytotoxic effects on human neoplastic cell lines

O. Sedláková, J. Sedlák, Ľ. Hunáková, J. Jakubíková, J. Duraj, M. Šulíková, J. Chovancová, B. Chorváth

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

Angiostatic substance TNP-470 displayed moderate cytotoxicity towards human leukemia HL-60, HL-60/ADR, HL-60/VCR and myeloma ARH77 cell lines with IC50 in the range 5-10 microM of concentrations and slightly higher IC50 for myeloma cell line U266. IC50 for ovarian CH-1, A2780 and A2780/ADR cell lines was in the range 10-15 microM with the exception of platinum-resistant SKOV3 cell line (more than 40 microM ). The IC50 values for MDA-MB-231 and MCF-7 breast carcinoma cell lines were 15 and 25 microM, respectively. In human hemopoietic neoplastic cell lines examined, TNP-470 induced the appearance of subpopulation with sub-G0 DNA content, suggesting the apoptosis-inducing potential of TNP-470 in these cells. No TNP-470-induced drug uptake modulation in drug-resistant leukemia cell line HL-60/VCR was observed. TNP-470 induced accumulation of cells in G0/G1 phase of cell cycle. There was no TNP-470-induced inhibition of MMP collagenase activity or MMP (MMP2 and MMP9) production in the human fibrosarcoma cells HT1080 in vitro.

Key words: TNP 470, angiogenesis inhibitor, cytotoxic activity in vitro, apoptosis, cell cycle alterations, human leukemia, myeloma and carcinoma cell lines, MMP2, MMP9, collagen zymography.
NEOPLASMA, 46, 5, 1999, 283-289

Characterization of APC exon 15 germ-line mutation in FAP family with severe phenotype showing extracolonic symptoms

T. Kirchhoff, Ľ. Kulcsár, M. Tomka, V. Števurková, V. Zajac

Cancer Research Institute, Slovak Academy of Sciences, 833 91 Bratislava, Slovak Republic;
National Cancer Institute, Bratislava, Slovak Republic

The adenomatous polyposis coli (APC) gene plays a crucial role in colorectal carcinogenesis. Germ-line mutations of APC gene give rise to familial adenomatous polyposis coli (FAP) - autosomal dominant syndrome manifesting hundreds to thousands of colorectal polyps, if untreated with malignant progression. We have used the techniques of heteroduplex analysis (HDA), protein truncation test (PTT), single strand conformation polymorphism (SSCP) and DNA sequencing for the identification and detailed positional analysis of mutations in FAP family with the expressive phenotype characterized by polyposis and extracolonic lesions. Detailed analysis revealed a 5bp deletion in a mutation cluster region (MCR) in exon 15 of APC gene in codon 1308. Two screened members of the FAP family exhibited this novel mutation.

Key words: Familial adenomatous polyposis (FAP), APC gene, heteroduplex analysis (HDA), protein truncation test (PTT), single strand conformation polymorphism (SSCP).
NEOPLASMA, 46, 5, 1999, 290-294

A note on nucleoli in granulocytic precursors of the granulopoietic proliferating compartment in patients suffering from the chronic phase of chronic myeloid leukemia treated with two different drugs with different mode of action

K. Smetana, I. Jirásková, H. Klamová

Institute of Hematology and Blood Transfusion, 128 20 Prague 2, Czech Republic

The incidence of main nucleolar types in granulocytic precursors was studied in the granulopoietic proliferating compartment (GPC) of patients suffering from chronic phase of the chronic myeloid leukemia (CML) who were treated by the widely used therapy with two different drugs with different mode of action - hydroxyurea (HU) and interferon alpha (IFN-alpha). In comparison with IFN early stages of GPC, i.e. myeloblasts and promyelocytes in patients treated with HU possessed more frequently micronucleoli which are known to reflect the direct as well as indirect inhibition of the nucleolar biosynthetic activities. On the other hand, the incidence of micronucleoli in these cells of a small percentage of patients treated with IFN also reached the average values of these nucleoli which were noted in patients treated with HU.

Key words: Nucleoli, granulopoietic proliferating compartment, chronic myeloid leukemia, chronic phase, hydroxyurea, interferon.
NEOPLASMA, 46, 5, 1999, 295-298

Expression of CD26 and DPP IV in T-acute lymphoblastic leukemia: Comparison of immunocytochemistry with enzyme cytochemistry

M. Klobušická, O. Babušíková

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

The possible identity of dipeptidyl peptidase IV (DPP IV) enzymatic activity and CD26 antigen expression in phenotypically defined T-acute lymphoblastic leukemia cells (T-ALL) was examined. For comparative studies, the combination of immunocytochemistry and enzyme cytochemistry methods was used. The strong correlation between the CD26 antigen expression and DPP IV positivity in the majority of T-lymphoblasts in T-ALL patients was evident. No CD26 antigen was expressed on DPP IV negative T-cells. The variable CD4 and/or CD8 antigen expression, frequent CD5 and CD7 positivity and absence of surface membrane CD3 antigen were the characteristic immunophenotypic features of CD26/DPP IV positive T-lymphoblasts. Moreover, the clear CD71 and CD26/DPP IV coexpression suggested the association of CD26/DPP IV positive cells with proliferation. The immunophenotype of CD26/DPP IV positive T-lymphoblasts seems to be characteristic for the relative immature cell population. In addition, noteworthy was the slight disassociation between the very high CD26 antigen expression and moderate DPP IV activity in cells of some T-ALL patients. The possible existence of enzymatically inactive structures of CD26 antigen or inactive precursors of DPP IV detectable only by immunocytochemistry was discussed. Our study indicates that CD26 antigen expression is tended to identify cells with DPP IV enzymatic activity in T-ALL patients. The results provide some more information of CD26 antigen involvement in the pathology of leukemic cells via its DPP IV enzyme activity.

Key words: T-acute lymphoblastic leukemia, CD26 antigen, dipeptidyl peptidase IV, immunophenotype.
NEOPLASMA, 46, 5, 1999, 299-303

Comparison of antileukemic immunity against U937 cells in atopic asthmatics versus healthy controls

Y.J. Chen, S.D. Shyur, C.Y. Wang, K.Y. Huang, Y.K. Chao, P.G. Chen

Department of Radiation Oncology, Mackay Memorial Hospital Taipei, Taiwan;
Department of Pediatrics, Mackay Memorial Hospital Taipei, Taiwan;
Department of Medical Research, Mackay Memorial Hospital, Taipei, Taiwan, 10449;
Department of Chinese Martial Arts, Chinese Culture University, Taipei, Taiwan, R.O.C.

To assess the antitumor effects in atopic asthmatics versus healthy adults, we designed this study using in vitro mononuclear cells (MNC) culture as an immunity model with human leukemic U937 cells as the target. MNCs were collected from asthmatic subjects and healthy controls. Conditioned media from the MNC cultures (MNC-CM) were collected after stimulation with various concentrations of phytohemagglutinin (PHA). We treated U937 cells with these MNC-CMs, then assayed their proliferation and differentiation after 5 days of culture. At lower PHA doses (1.25 microg/ml), as well as in absence of PHA, the asthmatic MNC-CMs inhibited U937 cells growth to a slightly greater extent than did the MNC-CMs from controls. In contrast, when higher doses of PHA were used (5, 10 microg/ml), this growth-inhibiting effect was dramatically reversed. The dual effect of MNC-CM in these two groups was also shown in U937 cell differentiation assay, assessed as follows: morphological change by Liu?s staining, functional change by NBT reduction test and CD14 expression by flow cytometric detection. We suggest that the antileukemic effects of MNCs from asthmatic patients result from a slightly immunopotentiated status. This immunity may be dramatically reversed, however, after marked activation of MNCs.

Key words: Antileukemic immunity, asthmatics, U937.
NEOPLASMA, 46, 5, 1999, 304-308

Metastasis as the first sign of thyroid cancer

L. Pomorski, M. Bartos

Clinic of Endocrinological and General Surgery, Institute of Endocrinology, Medical University of Łódź, 93 513 Łódź, Poland

The aim of this paper is to review our experience with patients who presented with a metastasic tumor in the lymph nodes or other organs as the first sign of thyroid cancer.
In 1974-1998, 18 602 patients were operated on due to goitre. There were 975 (5.2%) patients with thyroid malignant neoplasms. The group comprised 449 (46.1%) patients with papillary carcinoma, 309 (31.7%) with follicular carcinoma, 54 (5.5%) with medullary carcinoma, 106 (10.9%) with anaplastic carcinoma, and 57 (5.8%) with other types of thyroid malignant neoplasms.
Out of these 975 patients, thyroid cancer was diagnosed on the basis of the detection of a metastatic tumor in 26 (2.7%) patients. In 16 (61.5%) of these patients the metastatic tumor was located in the regional lymph nodes. In 10 (38.5%) patients distant metastasis beyond the regional lymph nodes was the first sign of thyroid cancer. In 5 (50%) patients metastasis was located in the bones, in 2 (20%) in the lung, in 1 (10%) in the heart, in 1 (10%) in the buttock, and in 1 (10%) in a central neck cyst. Metastasis was the initial manifestation of thyroid cancer in 18 (4%) of 449 papillary carcinoma patients, in 6 of 309 (1.9%) follicular carcinoma patients, and in 2 (3.7%) of 54 medullary carcinoma patients. Lymph node metastasis was the first sign of thyroid cancer in 13 (2.9%) patients with papillary carcinoma, 1 (0.3%) patients with follicular carcinoma and in 2 (3.7%) medullary carcinoma patients, and distant metastasis in 5 (1.1%) patients with papillary carcinoma and in 5 (1.6%) patients with follicular carcinoma.
After the detection of the primary focus of thyroid cancer total thyroidectomy and modified neck dissection were performed in all patients. Differentiated thyroid carcinoma patients were treated complementarily with 131I and TSH suppressive doses of l-thyroxine, and medullary cancer patients with teleradiotherapy and substitutive doses of l-thyroxine.

Key words: Thyroid cancer, metastasis, first symptom.
NEOPLASMA, 46, 5, 1999, 309-312

Osteoblasts and osteoclasts in bone marrow smears of cancer patients

A. Gaja, Z. Churý, N. Hejlová, H. Fraňková, D. Macková

Masaryk Memorial Cancer Institute, Brno, Czech Republic

Osteoblasts and osteoclasts are the unique cells occurring in bone marrow smears in situations with high bone metabolic turnover (children, trauma, rachitis, Paget disease or tumors). The collection of 2706 sternal or iliac crest aspirates from patients with hematologic malignancies and solid tumors are presented. We demonstrated significantly higher positivity for osteoblasts-osteoclasts presentation in bone marrow smears for hematological malignancies (p < 0.05), solid tumors (p < 0.01), and especially breast cancer (p < 0.001). We found a significant association between osteoblast-osteoclast positivity and dissemination of breast cancer (p < 0.05). None of the breast cancer patients without signs of dissemination (X-ray, sonography or scintigraphy) had positivity for osteoblasts or osteoclasts. We suppose that the osteoblast-osteoclast positivity in bone marrow smears can serve as a cheap marker for breast cancer dissemination.

Key words: Osteoblast, osteoclast, cancer, bone marrow, cancer dissemination.
NEOPLASMA, 46, 5, 1999, 313-316

Clodronate in the management of bone metastases: A clinical study of 91 patients

K. Serkies, B. Jereczek-Fossa, A. Badzio, J. Jassem

Department of Oncology and Radiotherapy, Medical University of Gdańsk, Gdańsk, Poland;
Department of Radiotherapy, European Institute of Oncology, Milan, Italy

The aim of our study was to evaluate the efficacy of oral clodronate supplementing systemic therapy and/or palliative irradiation in 91 patients with painful bone metastases. Clodronate was administered at a daily dose of 1600-3200 mg for a median of 11 months (range 3-36 months). Partial or complete pain relief was achieved in 61 of 88 evaluable patients (69%). Response rate to clodronate in patients who additionally received palliative bone radiation was similar to that in patients who did not receive irradiation (68 and 71%, respectively). Eleven out of 12 bed-ridden patients with metastatic bone pain regained the ability of walking after the treatment with clodronate. Bone pain relief lasted from 1.5 to 36 months (mean 9.3 months). Clodronate was well tolerated in all but three cases (3%) in whom the treatment was discontinued due to intensive adverse gastrointestinal effects. In conclusion, we observed satisfactory symptomatic effect and low rate of adverse reactions in patients with metastatic bone lesions treated with oral clodronate. Further large controlled studies with thorough patient monitoring are warranted to evaluate the real benefit of clodronate, and to define its optimal scheduling.

Key words: Bone metastases, clodronate, palliative treatment, bisphosphonates.
NEOPLASMA, 46, 5, 1999, 317-322

Quantitative analysis of modes of invasion and lymph node metastases in oral squamous cell carcinoma

A M. Altemani, P. Guimarăes, K. Metze, L.S. Queiroz

Department of Pathology, FCM-Universidade Estadual de Campinas (UNICAMP), 13 083-970 Campinas,SP, Brazil

The mode of tumor invasion has been suggested to have a relationship to the occurrence of cervical metastasis and to prognosis in oral squamous cell carcinoma (OSCC). However, a tumor usually does not have a single mode of invasion, and the importance, if any, of the relative proportions of different modes for metastatic potential has not been studied. Forty two cases of OSCC resected with cervical lymph nodes were selected, 20 of which had nodal metastases and 22 which had not. The mode of invasion in the tumor-host interface was classified as: I - pushing borders, II - bands, III - thin cords, IV - single cells and analyzed in 20 consecutive medium power fields. Also studied were other morphological parameters: perineural and angiolymphatic invasion, tissue eosinophilia, mitosis and intensity of inflammatory infiltrate at the tumor-host interface. The majority of the cases (95.2%) showed two or more modes of invasion. Modes I, II and III occurred with similar frequency in cases with and without metastases. Mode II was the commonest and most extensive in both groups. No mode of invasion was significantly associated with metastases, independent of its extension. The other morphological parameters were neither significantly associated with cervical metastasis. In conclusion, OSCC usually shows two or more modes of tumor invasion if a large extension of tumor-host interface is analyzed. However, the relative proportions of the modes have no correlation with the metastatic potential.

Key words: Oral squamous cell carcinoma, metastasis, lymph node, invasion.
NEOPLASMA, 46, 5, 1999, 323-328