Objective:To investigate the effects of erlotinib on proliferation and autophagy in HCC827,A549 and H1299 human non-small cell lung cancer cells. Methods:Three human non-small cell lung cancer cell lines HCC827,A549,H1299 were cultured in vitro,and were divided into control group,DMSO group,and groups treated by erlotinib in different concentrations. CCK-8 method was used for detecting the proliferation inhibition. The expression levels of autophagy-related proteins,microtubule-associated protein light chain (LC3)and Beclin-1 Were detected by Western blot analysis and cell immunofluorescence method. Results:Erlotinib inhibited the proliferation of HCC827,A549 and H1299 cells,proliferation inhibition rate showed that HCC827> A549 >H1299,IC50 of HCC827,A549 and H1299 cells were 8.72 μmol/L,32.09 μmol/L and 87.46 μmol/L,respectively. Compared with the control groups,erlotinib enhanced expression of LC3-II and Beclin-1 significantly(P < 0.05). Erlotinib induced autophagy in a concentration dependent manner in HCC827,A549 and H1299 cells(P < 0.05). Conclusion: Erlotinib inhibited proliferation selectively and activated autophagy in non-small cell lung cancer cells.