Objective:To study and compare the gene expresse of Oct4 and Nanog in human pancreatic cancer stem cell. Methods: FCM was used to isolate pancreatic cancer stem cells of CD44+CD24+ and CD44+CD24+ESA+ phenotypes by a FACS Aria II in human cell line Panc-1. Immunofluorescence was used to detect the expression of Oct4 and Nanog gene in Panc-1 cell and two kinds of cancer stem cell-like cells. Results:Oct4 and Nanog genes have a higher expression in CD44+CD24+ESA+ cells and CD44+CD24+ cells than Panc-1 cells. Conclusion:Oct4 and Nanog gene perform a higher expression in pancreatic cancer stem cells than in Panc-1 cells.