It is well established that TP53 regulates the expression of many genes, but the regulation of expression of TP53 itself is poorly understood. Recently, it has been shown that there is a tissue-specific binding of nuclear proteins in the TP53 gene promoter. The aim of this study was to determine the nuclear proteins that bind to the TP53 promoter elements (between -104 and -458) in male breast cancer. The results of our study, using the electrophoretic mobility shift assay (EMSA) and Southwestern analysis, have showed: (1) nuclear proteins or factors other than p53 bind to the TP53 promoter; (2) the levels of at least four nuclear proteins vary between normal and tumour breast tissue; and (3) two newly discovered nuclear proteins bind to the TP53 promoter in tumour tissue but are absent in normal tissue. This differential binding of nuclear proteins to the TP53 gene promoter might play a critical role in TP53 transcription and cancer progression in male breast tumours.