The genes of Gekkonidae, a lizard, are known to be very similar to human genes. According to previous research, lizard extracts inhibit angiogenesis and show anticancer activity against various cancers. In this regard, this study assessed whether lizard tail extracts (LTE) cause anticancer activity against lung cancer in mouse and human lung cancer cell lines. The results showed that LTE exhibited anticancer activity against lung cancer in vitro and in vivo. In vitro, cell viability and proliferation decreased in two lung cancer cell lines, while annexin V and single-stranded DNA both increased, showing apoptotic activity caused by LTE. We also found that LTE induced apoptosis in a caspase-3/7 cascade-dependent manner and inhibited the phosphorylation of Akt by participating in the PI3k/Akt pathway. In vivo, LTE decreased tumor volume in LLC bearing mice. Our results demonstrate the potential of LTE as a natural-derived anticancer drug to overcome the chemotherapy side effects during cancer treatment and contribute to the discovery of candidate substances.