This study investigated whether long noncoding RNA interfering EIF3J antisense RNA1 (EIF3J-AS1) could affect lung cancer A549 cells as well as the role of microRNA-330-5p (miR-330-5p) during this process. To this end, quantitative real-time polymerase chain reaction was used to measure EIF3J-AS1 and miR-330-5p expression in 39 lung cancer cases. Small interfering RNA targeting EIF3J-AS1 (si-EIF3J-AS1), as well as the miR-330-5p inhibitor, was transfected into lung cancer A549 cells. The outcomes of cell proliferation, clone formation, migration, invasion, and E- and N-cadherin expression were analyzed using CCK-8 kit, clone formation experiment, Transwell method, and Western blot. The targeted binding between EIF3J-AS1 and miR-330-5p was explored using the luciferase experiment. The results showed higher EIF3J-AS1 expression but lower miR330-5p expression cancer tissues. Furthermore, interfering EIF3J-AS1 increased miR-330-5p and E-cadherin protein expression, leading to a reduction in the proliferation, clone formation, migration, invasion, and N-cadherin protein expression of lung cancer A549 cells. Meanwhile, Transfecting si-EIF3J-AS1 and the miR-330-5p inhibitor could increase the proliferation, clone formation, migration, invasion, and N-cadherin protein expression of lung cancer A549 cells, suggesting the targeted relationship of EIF3J-AS1 to miR-330-5p. In summary, EIF3J-AS1 was highly expressed in lung cancer tissues, and interfering EIF3J-AS1 inhibited the proliferation, migration, and invasion of A549 cells through negative regulation of miR-330-5p.