〔Abstract〕 Objective To explore the effect of silencing human epididymal protein 4(HE4) on the proliferation, invasion, apoptosis and JAK/STAT3 signaling pathway of colon cancer cells. Methods 124 patients with colon cancer and 40 healthy people were selected. Western blot and immunohistochemical staining were used to detect the expression of HE4 in colon cancer and adjacent tissues. Enzyme linked immunosorbent assay(ELISA) was used to detect the expression of HE4 in serum of patients with colon cancer and healthy people. Western blot was used to detect the expression of HE4 in colon cancer cell lines Caco-2, SW480, HT-29, HCT-116 and hcoepic. HT-29 cells were transfected with HE4 si-RNA or si-control respectively. Cell proliferation was detected by CCK-8 method, invasion ability was detected by Transwell assay, and apoptosis was detected by Annexin-V-FITC/PI flow cytometry. Western blot was used to detect the protein expression levels of JAK1, p-JAK1, JAK2, p-JAK2, STAT3 and p-STAT3. Results The relative expression of HE4 protein in colon cancer tissues was higher than that in adjacent tissues(P<0.05). The expression of HE4 was positive in colon cancer tissues, of which 78 cases(62.90%) were highly expressed, while only 24 cases(19.35%) were overexpressed in the adjacent tissues(P<0.05). The serum HE4 concentration in patients with colon cancer was higher than that in normal subjects(P<0.05). The relative expression of HE4 protein in Caco-2, SW480, HT-29 and HCT-116 cells was higher than that in hcoepic cells(allP<0.05). The expression level of HE4 was not related to gender, age and tumor size, but related to differentiation degree, TNM stage, lymph node metastasis and distant metastasis(P<0.05). After silencing HE4 gene, the proliferation and invasion ability of HT-29 cells decreased, while the apoptosis rate increased(P<0.05). Silencing HE4 gene had no effect on the total protein of JAK1, JAK2 and STAT3, but could down regulate the expression of p-JAK1, p-JAK2 and p-STAT3(allP<0.05). Conclusion Silencing HE4 can inhibit colon cancer cell proliferation, invasion, and apoptosis, and down-regulate the expression of related proteins in the JAK/STAT3 signaling pathway.