Objective To observe the expression of Galectin-3 and Galectin-1 in pancreatic cancer and explore the relationship between the expression and pathological grading. Methods Forty specimens of pancreatic carcinoma tissue and thirty-one specimens of normal pancreas tissue were selected, which were confirmed by surgical resection and pathology from 2002 to 2009. The expression of Galectin-3 mRNA and Galectin-1 mRNA in pancreatic cancer cell lines SW1990, PANC-1 and ASPC -1 was detected by means of reverse transcriptase-polymerase chain reaction; the expression of Galectin-3 protein and Galectin-1 protein in SW1990, PANC-1 and ASPC-1 was detected by means of immunocytochemistry; the expression of Galectin-3 protein and Galectin-1 protein in pancreatic cancer and normal pancreatic tissue was detected by means of immunohistochemistry. Results In SW1990, PANC-1 and ASPC-1, Galectin-3 mRNA signal and protein were detected, but no Galectin-1 mRNA signal or protein was detected. There was no expression of Galectin-3 protein or Galectin-1 protein in the 31 specimens of normal pancrease tissue, while there were Galectin-3 protein and Galectin-1 protein expressed in the 40 specimens of pancreatic cancer tissue. In the 40 specimens of pancreatic cancer tissue, the expression of Galectin-3 protein was observed in pancreatic cancer cells, but not in fibroblasts or matrix cells around the cancer mass; while the expression of Galectin-1 protein was observed in fibroblasts and matrix cells around the cancer mass, but not observed in pancreatic cancer cells. There was no significant association between the expression of Galectin-3 protein in pancreatic cancer and pathological grading (P>0.05); while the expression of Galectin-1 protein in pancreatic cancer was related to the pathological grading, and the expression of Galectin-1 protein was significant higher in poorly differentiated tumors than that in well/moderately differentiated tumors (P<0.05). Conclusions Galectin-3 or Galectin-1 is not expressed in normal pancreases; Galectin-3 is expressed in pancreatic cancer cells; Galectin-1 is expressed in fibroblasts and matrix cells around the cancer mass. The expression of Galectin-1 is related with the differentiation of pancreatic cancer.