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Epigenetic silencing through promoter hypermethylation is an important hallmark for the inactivation of tumor-related genes in carcinogenesis. Here we identified the  ATP-binding cassette sub-family B member 4 (ABCB4)  as a novel epigenetically silenced target gene. We investigated the epigenetic regulation of  ABCB4  in 26 human lung, breast, skin, liver, head and neck cancer cells lines and in primary cancers by methylation and expression analysis. Hypermethylation of the  ABCB4  CpG island promoter occurred in 16 out of 26 (62%) human cancer cell lines. Aberrant methylation of  ABCB4  was also revealed in 39% of primary lung cancer and in 20% of head and neck cancer tissues. In 37% of primary lung cancer samples, ABCB4 expression was absent. For breast cancer a significant hypermethylation occurred in tumor tissues (41%) compared to matching normal samples (0%, p = 0.002). Silencing of  ABCB4  was reversed by 5-aza-2'-deoxycytidine and zebularine treatments leading to its reexpression in cancer cells. Overexpression of ABCB4 significantly suppressed colony formation and proliferation of lung cancer cells. Hypermethylation of  Abcb4  occurred also in murine cancer, but was not found in normal tissues. Our findings suggest that  ABCB4  is a frequently silenced gene in different cancers and it may act tumor suppressivly in lung cancer.

ABCB4 is frequently epigenetically silenced in human cancers and inhibits tumor growth