Objective To investigate the function of cAMP response element binding protein (CREB) and extracellular signal-regulated kinase (ERK1/2) in osteoclast differentiation mediated by Ca2+/calmodulin-dependent kinase Ⅱ (CaMKⅡ)δ, and elucidate the molecular mechanism thereof. Methods CaMKⅡδ RNA interference lentivirus vector was constructed and mouse RAW264.7 cells were transfected with the virus to determine the interference efficiency. After virus transfection, RAW264.7 cells were treated with 50ng/ml receptor activator of nuclear factor κB ligand (RANKL) and the phosphorylation levels of CREB and ERK1/2 were detected at different time points. The cells were also treated with PD98059, an ERK1/2 inhibitor, to determine the effect of ERK1/2 signal blocking on the expression of nuclear factor activated T-cells cytoplasmic 1 (NFATc1) and osteoclast differentiation. Results Interference efficiency of recombinant CaMKⅡδ virus vector was 77.2% at mRNA level and 70.2% at protein level. CaMKⅡδ RNA interference significantly suppressed phosphorylation of CREB and ERK1/2, and the levels of p-CREB and p-ERK1/2 were down-regulated by 21%-55% and 55%-64%, respectively. ERK1/2 inhibitor significantly down-regulated the protein expression of NFATc1, and the number of osteoclast, the number and size of bone resorption lacunae decreased by 39.3%, 50.0% and 52.3%, respectively. Conclusion CaMKⅡδ RNA interference may significantly suppress the phosphorylation of CREB and ERK1/2, and CREB and ERK1/2 have mediated the CaMKⅡδ-induced osteoclast differentiation. DOI: 10.11855/j.issn.0577-7402.2018.02.01