Objective: To investigate the potential role of p62dok in lipid-induced insulin resistance in adipocytes. Methods: p62dok expression and insulin signaling transduction were investigated in white fat tissues from mice treated with high-fat-diet (HFD). 3T3-L1 adipocytes were induced to insulin resistance by free fatty acid (FFA). Gene silencing of p62dok was used to study the effects of p62dok levels on insulin signaling transduction. Western blot was performed to detect protein levels and its phosphorylation statue. Results: The increased p62dok levels was found in white fat tissues from HFD-treated mice and in 3T3-L1 adipocytes treated with FFA, meanwhile insulin-induced phosphorylation of AKT was shown to decrease. The specific knock-down of p62dok in 3T3-L1 adipocytes treated with FFA resulted in increased insulin-induced phosphorylation of AKT, which was inhibited by Ras inhibitor Manumycin A and PI3K inhibitor LY294002, but not by MEK inhibitor U0126. Tyrosine-phosphorylation of IRS-1 did not show any changes in p62dok knock-down adipocytes treated with FFA. Conclusion: Our data suggest that lipid-induced upregulation of p62dok results in insulin resistance by inhibiting Ras/PI3K/AKT pathway in adipocytes.