DEPARTMENT OF GENETIC ECOTOXICOLOGY

Radim J. Šrám, M.D., D.Sc.
e-mail: sram@biomed.cas.cz
tel: +420241062596

Laboratory of Molecular Epidemiology

Head: Radim J. Šrám, M.D., D.Sc.
e-mail: sram@biomed.cas.cz
tel: +420241062596

Laboratory of Genetic Toxicology

Head: Jan Topinka, D.Sc.
e-mail: jtopinka@biomed.cas.cz
tel: +420241062675

Laboratory of Genomics

Head: Radim J. Šrám, M.D., D.Sc.
e-mail: sram@biomed.cas.cz
tel: +420241062596

Scientists:

Olena Beskid, Ph.D.
Miroslav Dostál, M.D., D.Sc.
Božena Novotná, Ph.D.
Pavel Rössner, M.D., D.Sc.
Pavel Rössner, Jr., Ph.D.
Jana Schmuczerová, Ph.D.

Ph.D. Students:

Mgr. Andrea Rössnerová
Oksana Sevastyanova, M.D.
Ing. Vlasta Švecová
Mgr. Yana Bagryantseva

Technical Assistants:

Dr. Irena Chvátalová
Dr. Eva Dejmková
Lenka Dostálová
Jana Herlová
Ing. Zdenka Lnenicková
Mgr. Alena Milcová
Mgr. Zuzana Nováková
Ing. Ivo Solanský
Olga Štveráková
Jolana Vaňková

Genetic damage caused by genotoxic and carcinogenic compounds

The research of the Department of Genetic Ecotoxicology (DGE) is concerned with the genetic damage caused by genotoxic and carcinogenic compounds such as polycyclic aromatic hydrocarbons (PAHs), alkenes, etc. The effects of genotoxic xenobiotics are studied in cell cultures, such as human hepatoma cell line HepG2 or human diploid lung fibroblasts, as well as in vivo by carrying out human transitional molecular epidemiologic studies and observational epidemiologic studies. The research is based on the application of advanced scientific technologies such as the 32P-postlabelling assay for DNA adduct analysis in lymphocytes and placentas, HPLC analysis of 32P-postlabeled DNA adducts, the expression of p53 and p21WAF1 proteins in plasma and lymphocytes, the comet assay to assess DNA damage in lymphocytes, cytogenetic analysis of chromosomal aberrations by conventional method as well as by fluorescence in situ hybridization (FISH), radioimmunological methods, flow cytometry for the phenotyping of lymphocyte subpopulations and methods for the analysis of genetic polymorphisms CYP1A1, GSTM1, GSTT1, GSTP1, NAT2, EPHX, XPD, XRCC1, hOGG1, p53, MS, MTHFR and APOE (PCR, RLFP techniques).
The DGE is a joint venture of the Health Institute of Central Bohemia and the Institute of Experimental Medicine AS CR. The DGE has been responsible for the scientific management of the Program "AIR POLLUTION AND HEALTH", which consists of several projects studying the impact of environmental pollution on the health of the population in Prague, as well as in the Northern and Southern Bohemia.
Current research at the Department focuses on the following main topics:

• Molecular epidemiology studies: biomarkers of exposure to mutagens and carcinogens
• Pregnancy outcome
• Polluted environment and health of children
• In vitro studies on genotoxicity of complex mixtures
• Oxidative damage of biomolecules
• Environmental pollutants and molecular mechanisms of prostate cancer
• Instability of DNA in patients with myelodysplastic syndromes (MDS)

Our studies suggest that carcinogenic polycyclic hydrocarbons bound on the respirable particles in ambient air are responsible for various adverse effects in general populations.

Fig. 1: Multistage process of chemical carcinogenesis

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Multiple steps from external and internal exposure through biologically effective dose and early genotoxic effects to changed structure and function are necessary to develop cancer resulting from carcinogen exposure. Moreover, other factors such as metabolic genotypes and phenotypes, DNA repair capacity and immune functions modulate the effect of genotoxic compounds in humans. To study most of these processes is the major aim of the Department of Genetic Ecotoxicology.

Fig. 2: Detection of DNA adducts by 32P-postlabelling

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Covalent binding of  genotoxic compounds and/or their metabolites is detected by 32P-postlabelling – ultrasensitive method of detection of DNA adducts. The method is used to detect DNA-reactivity of various chemicals and complex mixtures in vitro (acellular systems and cell cultures) as well as in animal and human tissues. Some DNA adducts might be fixed as gene mutations as a next step in multistage process of chemical carcinogenesis. DNA adducts in various human populations exposed to environmental mutagens and carcinogens helps to assess internal exposure for molecular epidemiology studies.

Fig. 3: Oxidative stress in cells

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Various methods may be used to detect oxidative stress in cells. We detect oxidative stress in lipids by measurement of  F2-isoprostanes (lipid peroxidation), in proteins by measurement of carbonyl groups and in DNA by detection of 8-oxo-dG by ELISA methods.

Fig. 4: Reciprocal translocation between chromosome 1 and 4 as detected by FISH

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Fluorescence in situ hybridisation (the whole chromosome painting of #1 and #4) is valuable tool to study chromosomal aberrations caused by various mutagens and carcinogens in the envioronment. The method is more sensitive compared to conventional cytogenetic analysis and it detects more serious chromosomal aberrations such as translocations.

Name of the grant	Grant agency	Number of the grant	Principal investigator	Duration of the grant
Impact of environmental and genetic factors on men fertility	GA AVČR	1QS500390506	Dr. Šrám	2005-08
Environmental factors and health	MŽP	VaV SL/5/160/05	Dr. Šrám	2005-07
Oxidative damage of DNA in bone marrow cells of patients with myelodysplastic syndromes - subtype refractory anemia with or without ring sideroblasts	IGA MZ	NR8265-3/2005	Dr. Novotná	2005-07
Application of toxicogenomics to study mechanisms of the impact of environmental pollutants on human health (ENVIRONGEN)	MŠMT ČR	2B06088	Dr. Šrám	2006-11
Modulation of anti-polycyclic aromatic hydrocarbon antibody level in relation to smoking and lung diseases (cancer/non-cancer); and the increase of polycyclic aromatic hydrocarbon resistance of organism by immunization	MŠMT ČR	2B06150	Dr. Topinka	2006-11
Integrated assessment of health risk of environmental stressors in Europe (INTARESE)	EC	018385-2	Dr. Šrám	2006-09
Assessing the risks of environmental stressors: contribution to development of integrating methodology (ENVIRISK)	EC	FP6-044232	Dr. Šrám	2007-09
Activation of pig oocytes using nitric oxide donors	GAČR	523/06/0295	Dr. Novotná	2006-08
The role of environmental pollutants in mechanisms regulating prostate carcinoma	GAČR	310/07/0961	Dr. Topinka	2007-10

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