CHARACTERIZATION OF mouse TESTICULAR TUMORS FROM formalin-fixed paraffin-embedded (FFPE) TISSUE BY PROTEIN MICROARRAYS

SILVINA QUINTANA; MARCELA VENARA; RODOLFO REY; CHRISTINA SCHOTT; MÓNICA VAZQUEZ-LEVIN; HÉCTOR CHEMES; KARL-FRIEDRICH BECKER

Barcelona, España

Congreso; IX International Congress of Andrology; 2009

International society of Andrology

Transgenic mice bearing a construct in which SV40 T expression is directed by the Antimullerian Hormone (AMH) promoter develop testicular tumors in adult life. Our aim was to extract immunoreactive proteins from FFPE mouse testicular tissue for protein array analysis. We studied the expression of EGFR, Cyclin D1, Cyclin D3, AKT, p-AKT, ERK y  p-ERK in protein lysates of 3 testis with normal and tumoral tissue (Whole tissue: WT) and in protein lysates of tumoral microdissected areas (TMA) of 7 different testis. After deparaffination, tissues were manually dissected from the slides and transferred into extraction buffer. From every protein lysate two dilutions (1:2, 1:4) and a negative control (only buffer) were prepared and applied onto a nitrocellulose coated glass slide. Relative expression levels of proteins were calculated by normalizing to total protein. AKT, p-AKT, ERK , p-ERK , EGFR and Cyclin D3 expression levels were higher in TMA in comparison with  WT (1,73 ± 0,61 vs. 0,95± 0,28; 1,70 ± 0,55 vs. 0,43± 0,12;  1,47 ± 0,52 vs. 0,69± 0,29 ; 3,72 ± 1,43 vs. 0,96 ± 0,32; 1,17± 0,37 vs 0,36± 0,13 and  1,18± 0,47 vs 0,35 ± 0,11  p< 0,01). No differences were found in Cyclin D1 expression between TMA and WT. These results demonstrate that it is possible to study protein expression levels from FFPE tissue by means of protein arrays. The higher levels of expression in tumoral tissue enriched areas (TMA) confirm an important activation of cellular proliferation pathways in tumoral tissues.