Objective: The aim of this study is to explore how quercetin interacts with pancreatic cancer stem-like cells and the mechanism underlying the effective quercetin-mediated suppression.Methods: A model of pancreatic cancer stem-like cells was generated by using a sphere formation culture system.A comparative analysis was performed between the parent cells and pancreatic cancer stem-like cells with a related treatment strategy focusing on cancer stem cell (CSC) properties and drug-resistance-related mechanisms in vitro.Results: Our data show that pancreatic cancer stem-like cells have greater resistance to gemcitabine and stronger CSC properties compared with the paint cells.In contrast to the pancreatic cancer stem-like cells, overexpression of β-catenin was observed in the parent cells.Quercetin suppressed proliferation, invasion and self-renewal capacity, and CSC surface markers expression, with alterations of β-catenin in pancreatic cancer stemlike cells.The combination of quercetin and gemcitabine can reduce tumor growth and decrease drug resistance in pancreatic cancer.Conclusions: β-Catenin plays an important role in maintenance and progression of pancreatic cancer.Targeting β-catenin using quercetin combined with gemcitabine may be a treatment strategy to improve prognosis in patients with pancreatic cancer.