VGF expression vector (pCMV6-AC–VGF–GFP, OriGene Technologies, Inc., Rockville, MD), as well as the empty vector (pCMV6-AC-GFP, OriGene Technologies, Inc.)were transfected into J82 and SCaBER bladder cancer cell lines that have fully methylated VGF promoter of our analyzed region and VGF silenced. The gene discussed is ASAH1; the disease is urinary bladder cancer.