Objective To establish a novel experimental neural invasion model with humanpancreatic cancercell line MIA PaCa-Ⅱand explore the biologicalcharacters. Methods The tumor mass which was formed by injecting humanpancreatic cancercell line MIA PaCa-Ⅱsubcutaneously was orthotopically transplanted into pancreas of nude mice. Morphological and biological features, metastasis and nerve invasion of the transplanted tumor were studied after 4, 6 and 8 weeks. Expression and distribution of K-ras, C-erbB2, COX-2, PSCA, p53 and DPC4 were detected in MIA PaCa-Ⅱ cell, with SABC immunohistochemistry. Results The successful rate of pancreatic cancer orthotopic implantation was all 100 % after 4, 6, and 8 weeks. The rate of nerve invasion was 50 %, 80 % and 60 % after 4, 6, and 8 weeks. Expression of K-ras, C-erbB2, COX-2 and PSCA were higher in pancreatic cancercells than in normal pancreascells. However, p53 and DPC4 expression were lower than normal. Conclusion Neural invasion model with humanpancreatic cancercell line MIA PaCa-Ⅱ orthotopic implantation tumor is successfully established in nude mice. The best time for exploring perinerval invasion is the sixth weeks. Every index studied may play important roles in the development of pancreatic cancer.