Mcl-1 (myeloid cell leukemia 1) is a prosurvival member of the Bcl-2 family and plays a critical role in cell survival by suppressing apoptosis through inhibiting the activity of proapoptotic proteins. It has been reported that Mcl-1 is frequently overexpressed in lung cancer. However, the exact molecular mechanism underlying Mcl-1 elevation in lung cancer is largely unknown. Here, we reported that Mcl-1 protein levels inversely correlate with FBXO4 expression, but not other F-box proteins examined, in lung cancer cell lines and lung cancer patient samples. Mechanically, FBXO4 is the E3 ubiquitin ligase to interact with and promote Mcl-1 ubiquitination and degradation. As a result, knockdown of Fbxo4 dramatically elevates Mcl-1 protein levels and increases cell survival and resistance to chemotherapeutic drugs, whereas ectopic expression of FBXO4 displays opposite phenotypes. Therefore, our study suggests that the protein stability of Mcl-1 is governed by FBXO4, which plays an important role in cell survival and chemotherapy for lung cancer.