IL-6, a pleiotropic cytokine, has been investigated for its role in regulating autophagy. Yet, its mechanism of action remains unclear. Here, we show that IL-6 exerted anti-autophagic effects on U937 cells through the STAT3 signaling pathway in vitro. The addition of IL-6 to starved U937 cells significantly activated the phosphorylation level of STAT3 (p-STAT3) at Tyr705 and reduced the protein levels of microtubule-associated protein 1 light chain 3 of type II (LC3-II) and Beclin 1. By immunoblotting, we also observed a positive correlation between the p-STAT3 level and Bcl-2 level. Furthermore, treatment with a STAT3 inhibitor, LLL12, or overexpression of a mutant form, STAT3Y705F, reversed the inhibitory effect of IL-6 on autophagy. Knockdown of Beclin 1 or Atg14 by siRNA and over-expression of Beclin 1 indicated the involvement of class III PI3K complex in IL-6-mediated inhibition of autophagy. Taken together, these data indicate that IL-6 inhibits starvation-induced autophagy and that p-STAT3 mediates the signal transduction from IL-6 to downstream proteins including Bcl-2 and Beclin1.