Cellular senescence is a state of irreversible growth arrest closely correlated with aging and may contribute to age-related disorders, which can be induced by telomere shortening (replicative senescence), oncogene activation and DNA damage (premature senescence). In the present study, we show that salubrinal, a selective inhibitor of eIF2α dephosphorylation, attenuates premature cellular senescence induced by etoposide. Etoposide treated cells exhibited senescent morphology, growth arrest and positive staining for senescence-associated β-galactosidase. The induction of the senescent phenotype was inhibited by the treatment of salubrinal. This suppression was associated the inhibition of p16INK4α gene promoters activity and a decreased level of p21 WAF1/CIP1 mRNA. The anti-senescence effect of salubrinal was mediated by activation of eukaryotic translation initiation factor 2α (eIF2α), because (1)salubrinal induced phosphorylation of eIF2α, (2) functional knockdown of eIF2α by GADD34 overexpression reversed the effect of salubrinal on senescence. In addition, level of ROS detected with ROS-responsive fluorescent probe was induced by etoposide, but suppressed by the treatment of salubrinal. Etoposide-induced expression of c-fos, an oxidative stress marker, was also inhibited by salubrinal. These results suggest that salubrinal interferes with senescence signaling via activation of eIF2α and suppression of oxidative stress.