Traf-2 and Nck interacting kinase (TNIK) is one of the STE20/MAP4K family members implicated in carcinogenesis and progression of several human malignancies. However, its expression pattern and biological behavior in pancreatic carcinoma remains completely unclear. The present study is designed to investigate the clinical and prognostic value of TNIK in pancreatic carcinoma. TNIK mRNA and protein level was respectively detected by real-time quantitative RCR (qPCR) and Western blot in ten paired samples of pancreatic cancer. Immunohistochemical staining was also conducted to examine TNIK in the tissue microarray (TMA) consisting of 91 archived specimens of pancreatic cancer. The correlation between TNIK and prognosis was assessed by Kaplan–Meier curves and Cox regression. The mRNA and protein levels of TNIK in pancreatic cancer were both significantly higher than those in matched paratumor tissues. Immunohistochemistry analysis showed that TNIK was positively associated with pathologic T (P = 0.045) and TNM (P = 0.040) stage. In addition, The Kaplan–Meier survival curves indicated that patients with high expression of TNIK had a shorter overall survival (OS) and disease-free survival (DFS) than those with low expression. Our results demonstrated that TNIK might play a crucial role in pancreatic carcinogenesis and serve as a novel therapeutic target of pancreatic cancer.