Characterization of mouse testicular tumors from formalin-fixed paraffin-embedded (FFPE) tissues by protein microarrays.

QUINTANA S.; SCHOTT C.; VENARA M.; REY R.; DI CLEMENTE N.; PICARD J.; VAZQUEZ-LEVIN, M.H.; CHEMES, H.; BECKER, K.

Catalonia Palace of Congress. Barcelona, España.

Congreso; 9th 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 evaluate the expression of protein proliferation markers from FFPE mouse testicular tissues by protein microarray analysis. We studied the expression of EGFR, Cyclin D1, Cyclin D3, AKT, phospho-AKT, ERK and  phospho-ERK in protein lysates from 3 testes containing normal and tumor tissue (Whole tissue: WT) or microdissected areas containing only tumor tissue (MTT) from 7 different testes of 7-10 month old mice. After deparaffination, tissue sections were manually dissected from the slides and transferred into protein extraction buffer (Qiagen). Relative protein expression was calculated by normalizing to total protein. Statistically significant higher expression levels (p< 0.05) were detected in MTT compared to WT for the following markers: AKT: 1.73 ± 0.6 vs. 0.95 ± 0.3; phospho-AKT: 1.70 ± 0.6 vs. 0.43± 0.1;   ERK: 1.47 ± 0.5 vs. 0.69± 0.3; phospho-ERK: 3.72 ± 1.4 vs. 0.96 ± 0.3; EGFR: 1.17± 0.4 vs 0.36± 0.1 and Cyclin D3: 1.18± 0.5 vs 0.35 ± 0.1. No differences were found in Cyclin D1 expression between MTT and WT. These results demonstrate the feasibility of assessing protein expression levels from mouse FFPE testicular tissue by means of protein microarrays. The higher expression levels found for EGFR, Cyclin D3, AKT, phospho-AKT, ERK and phospho-ERK in tumor enriched areas indicate a remarkable activation of cellular proliferation pathways in tumor tissues.