Abstract <p>Non-small cell lung cancer (NSCLC) is one of the most lethal types of malignancies around the world. Previously, LINC00852 is a tumor-associated lncRNA which has been verified to participate in affecting the activity of NSCLC cells in our work. However, its underlying molecular mechanism still remains poorly documented. The aim of this work is to further explore the involved regulatory mechanism of LINC00852 on NSCLC cells. Three NSCLC cell lines including H1299, A549 and LTEP-a2 cells were selected. LINC00852 was knockdown through its specific silence molecular ASO. Cell autophagy was analyzed by transmission electron microscope observation, fluorescent adenovirus infection and Western blot assays, respectively. Cell activity was measured through proliferation and colony formation assays. Knockdown the expression of LINC00852 leads to the activation of cell autophagy in three NSCLC cells. Previously, LINC00852 has been proved to act as a ceRNA of miR-29a-3p which further targets LAMTOR1. In this work, miR-29a-3p overexpression or LAMTOR1 knockdown both significantly increased the number of autolysosomes and promoted autophagy flux in NSCLC cells. Strikingly, the proliferation and colony formation abilities of NSCLC cells could be inhibited after treatment of rapamycin, an autophagy activator of cells. The results have indicated that knockdown of LINC00852 could impair the activities of NSCLC cells by activating cell autophagy via the LINC00852/miR-29a-3p/LAMTOR1 axis, which provides scientific guidance for the regulatory mechanism of long non-coding RNA LINC00852 in NSCLC cells, and also lay a theoretical basis for the potential treatment approach of NSCLC.</p>