beta1,4-galactosyltransferase II (beta1,4GalT II) is one of the enzymes transferring galactose to the terminal N-acetylglucosamine of complex-type N-glycans. Previously, we have reported that beta1,4GalT II overexpression increased cisplatin-induced HeLa cell apoptosis. However, the mechanisms of its expression regulation have been rarely investigated. Here, we cloned the 1.8-kb 5'-flanking region of the beta1,4GalT II gene and analysed its promoter activity. The transcriptional activity and mRNA expression level of beta1,4GalT II were dramatically induced by p53 transcription factor in HeLa cells. In response to DNA damage agent adriamycin, the mRNA expression and promoter activity of beta1,4GalT II were significantly up-regulated and the binding of p53 to beta1,4GalT II promoter was obviously increased. Furthermore, decreasing the expression of beta1,4GalT II using RNA interference inhibited p53-mediated HeLa cell apoptosis induced by adriamycin. Collectively, these results suggested that beta1,4GalT II might serve as a target gene of p53 transcription factor during adriamycin-induced HeLa cell apoptosis, which elucidated a new mechanism of p53-mediated cell apoptosis.