Background and Aims The activation of NF-κB plays a crucial role in development and progression of various malignant tumors. Studies demonstrated abnormal expressions in tumor necrosis factor receptor-associated factor 6 (TRAF6) and proteins associated with PTEN/PI3K signaling pathway in malignant tumors. Therefore, this study investigated the expressions of molecules mentioned above and their relations in colorectal cancer cells.Methods In colorectal cancer HT29 and SW480 cells after treatment with Maraviroc (CCR5 inhibitor), MG132 (TRAF6 inhibitor), or NF-BAY (NF-κB inhibitor), the changes in the expressions of those proteins, as well as in proliferation, migration and invasion abilities were detected by Western blot, CCK-8 assay and Transwell assay, respectively.Results In the two types of colorectal cancer cells, the PI3K expression was decreased and the PTEN expression was increased (all P<0.05), while the expressions of TRAF6 and NF-κB showed no significant changes (all P>0.05) after inhibition of CCR5 protein; the expressions of PI3K and CCR5 were decreased and PTEN expression was increased (all P<0.05), while the NF-κB expression did not significantly change (both P>0.05) after inhibition of TRAF6 protein; the expressions of CCR5, TRAF6 and PI3K were decreased and PTEN expression was increased after inhibition of NF-κB expression (all P<0.05). The proliferation, migration and invasion abilities in the two types of colorectal cancer cells were significantly suppressed by the treatment of any of the three inhibitors (all P<0.05).Conclusion There is constitutive activation of the NF-κB in colorectal cancer cells, which may inhibit the activity of tumor suppressor molecule PTEN, and thereby lead to the increased activity of tumor-promoting molecule PI3K as well as its pathway through up-regulating the expressions of TRAF6 and CCR5.