Free fatty acids (FFA) are closely related to insulin resistance in which proteins such as insulin receptor substrate 1 (IRS-1), phosphatidylinositol 3 kinase (PI3K) and glucose transporter type 4 (GLUT4) are involved. Recent researches have shown that G-protein coupled receptor 120 (GPR120) is a receptor for medium and long chain FFA. This study aimed to evaluate the relationship between GPR120 and proteins related to the glucose metabolism. We used siRNA technique to down-regulate the GPR120 expression in 3T3-L1 cells before incubation with palmitic acid (PA), and evaluated the effect of GPR120 expression on the levels of IRS-1, PI3K and GLUT4 after PA induction. RT-PCR and western blot were performed to detect gene and protein levels in differentiated cells. RT-PCR result showed that GPR120 mRNA significantly increased in differentiated cells. GPR120-SiRNA transfection significantly down-regulated GPR120 gene and protein level in differentiated 3T3-L1 cells on day 5 after PA-induced and also decreased lipid droplets accumulated within the cells. SiRNA-mediated decreased of GRP120 was associated with significant reductions in gene and protein levels of IRS-1 and GLUT4. Our results showed that the expression level of GPR120 affected the expression of proteins related to the glucose metabolism, suggesting a contribution of GPR120 in the insulin resistance.