Marchenko M.I.1, Bezdenezhnykh N.A.2, Kudriavets Y.l.2
COMPARATIVE CHARACTERISTICS OF THE EFFECT OF HEAVY METAL COMPOUNDS ON HUMAN CELLS CULTIVATED IN VITRO
1Institute for Occupational Health of AMS of Ukraine, Kiev
2Institute of experimental pathology, oncology and radiobiology n. R.E. Kavetsky of NAN of Ukraine, Kiev
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In order to model the assessment of chemical toxicity of cells in vitro experiments have been conducted, using human cells of different origin (carcinomic alveolar basal epithelial cells and hepatocarcinoma cell line A-549, A-549IFN, Hep-G2). Two basic tests were used for comparative studies, where the number of viable cells was estimated in the culture — MTT (methyl tetrazolium test) and sulforhodamine B method. For the test, four samples of different salts of heavy metals — MnS04, CdS04, Pb acetate and HgCl2, were taken, being the most prevalent in the environment. It was established that two different methods for the analysis in vitro did not differ significantly from each other by sensitivity (according to the concentration 1C, resulting in 50 % loss of cells in vitro). However, for the purpose of monitoring of a greater number of chemical substances the sulforhodamine B method is more acceptable and not so expensive. It was confirmed that cells of different tissue origin dif¬fered by sensitivity to some xenobiotics. It is shown that modification of cells in a long-term exposure to interferon can increase cell sensitivity to salts of some metals. The results obtained in vitro correspond to the results of studies on acute tox¬icity of heavy metals in vivo tests.
Key words: basal cytotoxicity, IC50, cell cultures, heavy metals, MTT-test, sulforhodamine B method, interferon modified cells
- German A. et al. ECVAM Workshop 50: strategies to replace in vivo acute systemic toxicity testing - 2004.
- ICCVAM (2001) Report of the International Workshop on In Vitro Methods for Assessing Acute systemic Toxicity. ICCVAM-NICEATM workshop. Arlington, VA. USA.- 2000,- NIH Publication No 01-4499.- P.370
- Kundiyev YU.I., Trakhtenberg I.M. (2007), Khimicheskaya bezopasnost' v Ukraine. Yezhegodnyye chteniya, posvyashchennyye pamyati Ye.I. Goncharuka [Chemical safety in Ukraine. Annual readings dedicated to the memory of E.I. Goncharuk], Bogomolets National Medical University, Avicenna, Kyiv, Ukraine.
- Belyaeva Е.А. et al. Cd2+-promoted mitochondrial permeability transition: a comparison with other heavy metals // Acta Biochimica Polonica.- 2004 - V.51, No 2,- P.545-551.
- Duruible J.O., Ogwuegbu M.O.C., Egwurugwu J.N. Heavy metal pollution and human biotoxic effects // International Journal of Physical Sciences – 2007, V. 2, No 5.
- Scudiero D. et al. Evaluation of a Soluble Tetrasoliu/Formazan assay for cell growth and drug sensitivity in Culture using human and other cell lines // Cancer Res - 1988,- V.48 - P. 4827-4833
- Shekan P. et al. Evaluation of Colorimetric Protein and biomass Stains for Assaying Drug Effects Upon Human Tumor Cell lines // Proc. Amer. Assoc. Cancer Res., - 1989 - V.30 - P. 2436
- Liu D. A rapid biochemical test for measuring chemical toxicity to bacteria of single chemicals and mixture // Toxicity Screening Procedures Using Bacterial Systems, New York, 1981,-P. 175- 194
- Vorontsova A.L., Kudryavets YU.I. Zhil'chuk V.Ye. Interferony i ikh primeneniye v klinicheskoy onkologii // Zdorov'ye zhenshchiny,- 2003.- T.4, No 16,- S. 8-14.
- Satomi Н., Wang В., Fujisawa Н., Otsuka F. Interferon-beta from melanoma cells suppresses the proliferations of melanoma cells in an autocrine manner// Cytokine Interferon.-2002.-Apr 21. No 18(2).-P. 108-15.
- Kudryavets YU.Y. Interferon-alfa posylyuye rozvytok apoptozu, indukovanoho riznymy chynnykamy v pukhlynnykh klitynakh in vitro // Éksperym.onkolohyya,- 2001.- V.23, No 4,- P.267-73.
- Bezdyenyezhnykh H.O., Lukyanova N.YU., Lykhova A.A., Kudryavets Yu.Y., Doslidzhennya mekhanizmiv chutlyvosti klityn raku leheni lyudyny do kombinovanoho vplyvu khimiopreparativ i hipertermiyi v umovakh prolonhovanoyi diyi interferonu in vitro // Materialy IX Mizhn. konf. molodykh onkolohiv, Kyiv 2008.- S.20-21.
- Bezdenezhnykh N.A., Luk'yanova N.YU., Vorontsova A.L., Kudryavets Yu.I., Markery kletochnoy proliferatsii, differentsirovki i zlokachestvennosti pri dlitel'noy ekspozitsii kletok nemelkokletochnogo raka legkogo s interferonom in vitro // Materíali IX Mízhn. konf. molodikh onkologív, Kyiv, 2008.- S. 22-23.