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There is emerging evidence on the mechanisms of pancreatic cancer pain. Following the establishment of an orthotropic transplantation model of pancreatic cancer, microarray analysis was performed to identify changes in the expression levels of pain-associated genes in the spinal cord. A mouse model of pancreatic cancer-induced pain was established by implanting SW 1990 cells into the pancreases of female BALB/c-nu mice. The survival rate and body weight were measured following orthotropic transplantation. Gross anatomical techniques and hematoxylin and eosin staining were used to analyze the pancreatic tumor tissue. Multiple behavioral tests were also performed to assess pain-associated responses. Additionally, using samples from mice with or without observable pain, microarray analysis was performed to determine the gene expression profiles in the spinal cord dorsal horn. The survival rate of mice with pancreatic cancer was high during the initial 3 weeks post-surgery, although the body weight decreased progressively. Gross anatomical techniques demonstrated that the tumor size increased significantly following the surgery, and this result was confirmed by solid tumor masses in the pancreatic tissues of the mouse model. Observable pain behavioral responses were also examined in the pancreatic cancer model by measuring the mechanical threshold of the abdominal skin, hunching behavior and visceromotor responses. The profiles of 10 pain specific-associated genes in the spinal cord dorsal horn that accurately reflect the molecular pathological progression of disease were also identified. In conclusion, the present study has developed a novel animal model of pancreatic cancer pain in BALB/c-nu mice that resembles human pancreatic cancer pain, and the expression of pain-associated genes in the spinal cord dorsal horn has been profiled. The results of the present study may further the understanding of the molecular mechanisms that mediate pancreatic cancer pain.