I. Identification and characterisation of epigenetically altered tumor supressor genes by proteomic and genomic approches / II. Studies of genes regualted by MeCP2 union to its promoter regions by proteomic and genmomic approches

DNA methylation has an important impact on normal cell physiology, thus any defects in this mechanism may be related to the development of various diseases In this project we are interested in identifying epigeneticaliy modified genes, in general controlled by processes related to the DNA methylation, by means of a new strategy combining protomic and genomic analyses. First, the two Dimensional-Difference Gel Electrophoresis (2-DIGE) protein analyses of extracts obtained from HCT-116 wt and double knockout for DNMT1 and DNMT3b (DKO) cells revealed 34 proteins overexpressed in the condition of DNMTs depletion. From five genes with higher transcript lavels in DKO cells, comparing with HCT-116 wt. oniy AKR1B1, UCHLl and VIM are melhylated in HCT-116. As expected. the DNA methvlation 1s lost in DKO cells. The rneth,vl ation of VIM and UCHLl promoters in some cancer samples has already been repaired, thus further studies has been focused on AKRlBI. AKR1B1 expression due lo DNA methyiaton of promoter region seems to occur specilfically in the colon cancer cell Iines. which was confirmed in the DNA rnethylation status and expression analyses. performed on 32 different cancer cell lines (including colon, breast, lymphoma, leukemia, neuroblastoma, glioma and lung cancer cell Iines) as well as normal colon and normal lymphocytes samples. AKRIBI expression after treatments with DNA demethvlating agent (AZA) was rescued in 5 coloncancer cell lines (including genetic regulation of the candidate gene. The methylation status of the rest of the genes identified in proteomic analysis was checked by methylation specific PCR (MSP) experiment and all appeared to be unmethylated. The similar research has been done also bv means of Mecp2-null mouse model For 14 selected candidate genes the analyses of expression leveis, methylation Status and MeCP2 interaction with promoters are currently being performed.

Monitorizacion de los cambios de uso del suelo en la cabecera de cuenca de la ribera salada mediante fotografía aérea y S.I.G. (El Solsonès, Lleida, España)

Durante estos últimos 50 años ha tenido lugar un abandono de las masías y de los cultivos tradicionales en los Prepirineos catalanes. En este trabajo se han estudiado los cambios de usos del suelo desde los años 50 hasta la actualidad en una cuenca de 6577 ha situada en la cabecera de la Ribera Salada (El Solsonès). Para ello se han utilizado fotografías aéreas del año 1957 y del año 1992. Como base cartográfica sirvieron los ortofotomapas del año 1993. El análisis de los cambios de usos según diferentes variables (pendiente y orientación) y según diferentes enfoques (intensidad del cambio y uso final), se ha llevado a cabo mediante el programa de SIG ARC/INFO®. Se han establecido tendencias en la evolución de los usos según la pendiente y la orientación. Los resultados obtenidos muestran que los campos de cultivo son los que han disminuido más en superficie, dando lugar a pastos. Por otro lado, el bosque denso ha aumentado considerablemente su extensión (845 ha).