Amplification and overexpression of vinculin are associated with increased tumor cell proliferation and progression in advanced prostate cancer. Homo sapiens

We analyzed the 10q22 amplification in advanced prostate cancer and subjected the genes located in the common amplified region to an RNAi screen. We found vinculin as the most promising candidate gene and analyzed its protein expression on more than 400 prostate cancers by using the tissue micrarray (TMA) technology. We discovered a strong correlation between amplification of the vinculin gene and increased protein expression. Further, vinculin protein expression was strongly associated with increased tumor cell proliferation. Overall design: We isolated DNA from four 10q22 amplified cell lines (PC-3, MFM-223, SK-BR-3, ME-180). Genomic DNA was digested with the restriction enzymes RSA I / ALU I and then labeled with Cy5 using a Klenow-based commercial kit (Invitrogen). Each sample was hybridized with a pooled normal (46,XX) reference (Promega) to Agilent 244k CGH arrays. This experiment allowed us to define the common amplified region of the 10q22 amplification.