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We examined whether patterns of multigene methylation were simply biologic signatures of aggressive PENs or reflected sequential accumulation of methylation at multiple loci during tumor progression. Unfortunately, from the 14 PENs with synchronous hepatic metastases, we were only able to study metastatic tumors correlating to primary lesions in 4 cases. Although 1 of these neoplasms (sample 28-M) acquired methylation at 1 additional gene during metastasis, the other 3 cases demonstrated complete matching of the methylation profiles for the primary and metastatic tumors. From these data, we cannot conclude that gene methylation accumulates during tumor progression or even during metastatic spread. However, this consistency of methylation between primary and secondary neoplasms may have clinical application. The methylation status of individual or multiple genes could be used to follow tumor aggressiveness in response to surgical and/or medical therapy or to prove which primary tumor (if more than one) is responsible for any synchronous or metachronous metastasis. Also, specific methylation patterns of histopathologically normal tissues within the surgical specimen, including adjacent organs, lymph nodes, or distant sites, may be indicative of occult regional or systemic tumor spread. 1e1e36bf2d