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Purpose Defensin alpha 6 (DEFA6) is cationic short peptide with known functional activities in innate antimicrobial immunity. DEFA6 is also highly expressed in colorectal cancer tissue. The mechanism and function of DEFA6 have not been reported how to play a significant role in carcinogenesis and cancer progression. The aim of this study was to evaluate the protumorigenic functions of DEFA6 in the colorectal cancer cell line and the clinical significance of DEFA6 expression in colorectal cancer patients. Methods DEFA6 expression was investigated by immunohistochemistry in 151 cases of colorectal cancer tissue and the association of DEFA6 expression was correlated with patient's cancer charecteristics. Results Immunohistochemistry analysis showed that the DEFA6 protein was expressed higher in advanced cancer group (high T stage, patients with lymph node metastasis, patients with vascular invasion) than early cancer group (low T stage, patients without lymph node metastasis, patients without vascular invasion) (P=0.007/0.032/0.047). Conclusion As patient's survival usually depends on migration and invasion of the cancer cell, the high expression of DEFA6 in colorectal cancer cell is associated with patient's cancer charecteristics and could be a biomarker for colorectal cancer. The future study about characterization of DEFA6 will clarify its specific role in oncogenesis and its therapeutic potential.