The oncogene Akt1 plays a pivotal role in regulating growth factor-associated cell survival. The activity of Akt must be carefully regulated as it has been reported that an increase in Akt signaling is frequently associated with cancer. Phosphorylated Akt, as the constitutively activated form of Akt, has been observed in many kinds of cancer, but the clinical relevance of the (DNA) methylation profile of Akt1 in cancer is not well understood. This study aims to investigate the methylation level of Akt1 in bladder cancer tissues and their clinical significance. Methylation of the oncogene Akt1 transcriptional regulation region (TRR) was detected using bisulfite-specific (BSP) PCR-based sequencing analysis in cases of bladder cancer and the normal tissues, including 15 bladder cancer tissues, and five normal bladder tissues. BSP cloning-based sequencing analysis was also performed in selected cases. Clinicopathological data from the cancer patients were collected and analyzed. Analysis of Akt1 gene TRR methylation showed decreased methylation level in bladder cancer than normal. Methylation level of Akt1 has clinical relevance (P = 0.0043 by unpaired student's t test) with bladder cancer. Abnormal methylation of the Akt1 gene may be an early event during urocystic tumorigenesis and should be further evaluated as a tumorigenesis marker for early detection of bladder cancer.