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180 Abstracts / Toxicology L

mblyomin-X treatment (10 or 100 ng/10 �L) was topically appliedach 48 h in the absence or presence of B16F10 lineage, whichas injected into microcirculatory beds of the subcutaneous tissue.umbers of vessels and tumor growth were quantified in imagesbtained before and at 8th day after beginning of treatments. Aftericrocirculatory endothelial cell lineage (T-end lineage) conflu-

nce, a mechanic lesion in the culture was done by a cell scraper andmblyomin-X (10 or 100 ng/well) or/and VEGF (100 ng/mL) weredded to the wells. Cell migration was monitored at 12 h by theumber of cells migrating into the lesion and PGE2 and nitric oxideNO) were quantified in the supernatant. Results: Amblyomin-Xignificantly reduced the number of vessels in the subcutaneousicrocirculation (10 ng = 31.7% and 100 ng/10 �L = 42.7% in com-

arison to the first day of treatment) and the tumor growth. Onlyn the absence of VEGF, Amblyomin-X reduced endothelial cell

igration into the lesion focus during all period of time mon-tored, independently of PGE2 and NO secretion (10 ng = 16.4%nd 100 ng/10 �L = 23.1% in comparison to the control). Discussion:ocal application of Amblyomin-X reduces, in vivo, the num-er of microcirculatory vessels and tumor growth. Impairmentf endothelial cell migration may suggest an effect on new ves-els formation. Future investigations will be carried out to clarifyhe mechanisms involved in this process. Financial support: CAPES,APESP grant 08/57850-8.

oi:10.1016/j.toxlet.2010.03.612

203-023ffects of the environmental pesticide DDT and its metabolitesn the human breast cancer cell line MCF-7

. Pestana 1, D. Teixeira 1, A. Faria 1,2, V. Domingues 3, R.onteiro 1, C. Calhau 1

Department of Biochemistry (U38-FCT), Faculty of Medicine,niversity of Porto, Portugal, 2 Chemistry Investigation Centre (CIQ),aculty of Sciences, University of Porto, Portugal, 3 Requimte –nstituto Superior de Engenharia, Instituto Politécnico do Porto,ortugal

uman exposure to persistent organic pollutantsPOPs) is a certainty, even to long banned pesticidesike dichlorodiphenyltrichloroethane (DDT), and its

etabolites dichlorodiphenyldichloroethylene (DDE) andichlorodiphenyldichloroethane (DDD). POPs are known toe particularly toxic and have been associated with endocrine-isrupting effects in several mammals, including humans even atery low doses. The discovery that low concentrations, detectedn different human samples, may act on certain molecular mech-nisms is leading to a reassessment of metabolic POPs impact onuman beings.

In this regard, the effects of o,p′-DDT, p,p′-DDE and p,p′-DDDere assessed on a hormone-sensitive breast cancer cell line (MCF-

). For this purpose, proliferation and viability in addition to theirnvasive potential were assayed, in the presence (50–1000 nM) orbsence of the tested compounds. Proliferation was evaluated byethyl-3H-thymidine incorporation into DNA and viability trough

he lactate dehydrogenase assay. Invasion potential was assessedsing a commercial fluorometric cell invasion assay.

Cell proliferation and viability was not equally affected byhese compounds. A significant decrease in cell proliferation

as observed in the presence of o,p′-DDT (100–500 nM). In

he presence of DDT metabolites, p,p′-DDD, p,p′-DDE, prolifera-ion was not significantly affected for all tested concentrations50–1000 nM), excepting a significant reduction with 50 nM of

196S (2010) S37–S351

p,p′-DDD. Additionally, all compounds showed cytotoxicity withdifferent concentrations (50 and 500 nM p,p′-DDD; 500 and1000 nM p,p′-DDE; 100 and 250 nM o,p′-DDT).

Cell invasion was significantly induced by all tested compounds,in concentrations that were not able to decrease proliferation orincrease cytotoxicity.

Altogether, these results showed that these compounds werecytotoxic and able to induce cell invasion. Considering their knownestrogenic activity, further research is needed to clarify the signalpathways herein involved.

Financed by FCT (POCI, FEDER, Programa Comunitáriode Apoio, PTDC/QUI/65501/2006; SFRH/BD/46640/2008,SFRH/BD/64691/2009, SFRH/BD/28160/2006 andSFRH/BPD/40110/2007).

doi:10.1016/j.toxlet.2010.03.613

P203-024Increased breast cancer risk in women with environmentalexposure to pesticides

T. Parron 1, R. Alarcon 2, M.D.M. Requena 1, A. Hernandez 3

1 Delegacion Provincial de Salud de Almeria, Spain, 2 Universidad deAlmeria, Spain, 3 Universidad de Granada, Spain

Exposure to environmental pollutants has raised health concerns,especially in relation to its carcinogenicity effects. Pesticides havebeen considered as risk factors for breast cancer since these envi-ronmental chemicals exert estrogenic activity on in vitro and in vivomodels. This fact, together with the increasing incidence of breastcancer in Spain and the lack of a clear relationship between pesti-cide exposure and this disease led us to evaluate the role of thesecompounds as a risk factor for breast cancer in women Almeria.

We have performed a descriptive epidemiological study to col-lect data from all cases of breast cancer for the study period1998–2009 through the hospital minimum dataset from the centralreference Hospital of Almeria province. This province was classi-fied in two different geographical areas according to the numberof hectares devoted to intensive agriculture: high exposure areas(>1000 ha of greenhouses) and low exposure areas (<1000 ha).

A total number of 2661 cases of breast cancer were collectedin female population, of whom 2173 were observed in areas ofhigh pesticide exposure (rate of 81.07 per 100,000 population) and488 cases in low exposure areas (64.71 per 100,000 population).The population living in areas of high pesticide exposure was at anincreased risk for breast cancer as compared to that living in areasof low pesticide exposure (OR 1.25, 95% CI 1.13–1.38). In conclu-sion, this study supports the relationship between environmentalcontamination by pesticides and breast cancer. Nevertheless, thistype of study cannot be considered as a proof of causality.

doi:10.1016/j.toxlet.2010.03.614

P203-025A liver cancer study comparing two geographical areas withdifferent levels of pesticide use

T. Parron 1, R. Alarcon 2, M.D.M. Requena 1, A. Hernandez 3, A. Pla 3

1 Delegacion Provincial de Salud de Almeria, Spain, 2 Universidad deAlmeria, Spain, 3 Universidad de Granada, Spain

The relationship between pesticide exposure and liver cancer hasbeen reported in different studies although most of them are