Karyotype on human GSM-exposed amniocytes
Abstract
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Human amniocytes will be exposed for 24 h to GSM-900 in a wire-patch cell. 0h and 24 h post-exposure, karyotypes (R-banding) of the cells will be performed in order to determine the number of aneuploidies, the number of structural rearrangements and of DNA breaks. The micronuclei test will also be performed, in order to compare the results obtained by two different technics. A relation between SAR and effect will also be studied.
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Partners
Faculté de Médecine de Limoges – Laboratoire de Cytogénétique
http://www.unilim.fr/medecine/
Institut de recherche XLIM UMR CNRS 6172
http://www.xlim.fr/fr/ircom.jsp
Contact
Catherine YARDIN – Faculté de medecine de Limoges
Duration
24 months
Summary
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Our aim is to evaluate the genotoxic effects of the electromagnetic waves (EW) in vitro on fibroblastic human cells: amiocytes provided from the Department of Cytogenetics of the Limoges Hospital. Complete karyotypes of exposed cells will be performed and compared to sham cells. Cells will be exposed in a wire-patch (Laval et al., 2000). The test of micronuclei will also be used in order to see whether both technics give the same results.
This project results from a collaboration with the research institute XLIM (Limoges), particularly with a project named “electromagnetic waves and health”. This collaboration already led us to realise some works concerning EW and apoptosis. The studies were performed by a PhD student (V. Joubert) whose work is entitled “neuronal cells exposure to radiofrequencies: study of apoptosis”. The first results are published (Joubert et al., 2006; Joubert et al., 2007) and another study is submitted for publication.
For the present study, another PhD student specialised in human cytogenetics (S. Bourthoumieu) will specifically work on this subject. She will eventually be graduated during the year 2008-2009.
Until now, the works concerning the genotoxic effects of radiofrequencies gave opposite results and need further confirmation. A recent review of the literature (Vijayalaxmi et Obe, 2005) analysed the different results obtained and concluded that further studies are necessary using human cells, for example fibroblasts. As far as fibroblastic cultures are obtained after skin biopsy that is an invasive technique, we chose to work on amniocytes. These cells will be provided by the department of Cytogenetics after amniocentesis for fetal karyotype, and with consent of the patients. For the study the cells will be used once the diagnostic analysis will be performed.
The R-banded karyotype is a reliable technique to evaluate clastogenic and aneuploidogenic effects. The rates of chromosomal breaks, rearrangements and aneuploidies obtained on the GSM-900 (24 h, SAR at 2 W/Kg) exposed amniocytes will be compared to those of sham cells providing from the same amniotic fluid sample. 100 metaphases will be examined per condition (0 H and 24 H post-exposure). At least 5 amniotic fluid samples will be analysed (at least 2000 metaphases analysed). In order to interpretate the results a statistical analysis will be performed. Furthermore a same sample of amniotic fluid will be exposed to different SAR during a constant time (24 h) to determine a possible dose-effect.
To our knowledge, the complete karyotype has never been used before in this sort of study, because of the lack of experiment people. We have ministerial agreement for prenatal and postnatal cytogenetic analysis. In a second time, the micronuclei test will be used to verify the results by a second technique.