To investigate the mechanism of miR-144-3p in lung adenocarcinoma (LUAD), the expression le-vels of miR-144-3p and its downstream target gene in LUAD tissues were predicted based on multiple databases. Through bioinformatics methods and based on the related literature, E2F transcription factor 8 (E2F8) was identified as the target gene of miR-144-3p, and the dual-luciferase reporter gene assay was conducted to verify the target binding relationship between miR-144-3p and E2F8. The regulatory mecha-nisms of miR-144-3p and E2F8 in LUAD were explored by real-time quantitative PCR (qRT-PCR), Western-blot, Cell Counting Kit-8 (CCK-8) assay, colony formation assay, invasion assay and flow cytometry. The re-sults showed that miR-144-3p expression was significantly low in LUAD tissues and cells, while E2F8 ex-pression was significantly high. There was a targeting relationship between miR-144-3p and E2F8. Overex-pression of miR-144-3p or knockdown of E2F8 inhibited the proliferation and invasion of LUAD cells and arrested the cell cycle at G1 phase. In summary, miR-144-3p could inhibit the proliferation and invasion of LUAD cells and arrest the cell cycle at G1 phase by negatively regulating E2F8 expression, thereby inhibi-ting the malignant progression of LUAD cells. The results may be helpful for developing new methods in lung cancer detection and treatment.