The human mucin gene MUC4 is overexpressed in pancreatic cancer and cancer cell lines, while remaining undetectable in the normal pancreas, indicating its important role in pancreatic cancer pathogenesis. The molecular mechanisms involved in the regulation of MUC4 gene expression are not fully understood. In this report, we used pancreatic cancer cell line (Bxpc-3) to explore the potential transcription factors regulating MUC4 transcriptional activity. Through promoter screening, overexpressing methods and luciferase reporter studies, we found that transcription factors CREB, Ets-1, Elk-1 and STAT1 can positively regulate MUC4 expression at the promoter and mRNA level. Our findings will be helpful for better understanding the transcriptional regulation of MUC4 in pancreatic cancer cells and identifying key biologically relevant factors that may account for its aberrant expression in pancreatic cancer.