Objective To investigate the effect of Genistein on Akt/NF-κB pathway mediated interleukin-8 (IL-8) secretion in human colon carcinoma HT29. Methods Cells were treated with LPS plus hIFN- γ.Cellular proliferation of HT29 cells was measured by thiazolyl blue tetrazolium bromide (MTT) test. Expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), IL-8 and interleukin-10 (IL-10) was identified by quantitative real timepolymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). Western blot method was performed to detect the expression of p-Akt (S473), Akt, p-p65 and p65. Results Genistein (200 μmol/L) had no obvious inhibitory effect on HT29 cell proliferation. ELISA and qRT-PCR confirmed reduction of LPS+hIFN-γ induced IL-8 in Genistein treated HT29. Western blot analysis indicated that Genistein inhibited the activation of Akt/NF-κB induced by LPS plus hIFN-γ. Conclusion Genistein inhibits expression of IL-8 through inhibiting the activation of Akt/NF-κB pathway in HT29 cell line.