Abstract: Objective To investigate the effect of Anlotinib on proliferation and apoptosis in non-small cell lung cancer(NSCLC) cells and its molecular mechanism. Methods Non-small cell lung cancer cell lines A549 and H1299 were incubated with Anlotinib, miR-16-5p agonist and/or PD-1 overexpression vector respectively. CCK-8 assay and EDU assay were applied to detect the proliferation. Flow cytometry was performed to detect the cell apoptosis. The relative expression of miR-16-5 p in A549 and H1299 was detected by real-time quantitative polymerase chain reaction(RT-qPCR). The relative protein expression of PD-1 in A549 and H1299 was detected by Western blot assay. The interaction between miR-16-5p and PD-1 was determined by dual luciferase reporter assay. Finally, A549 cell xenograft model was established to assess the effect of Anlotinib on tumor growth in vivo. Results Anlotinib significantly increased miR-16-5p expression and decreased PD-1 expression in A549 cells and H1299 cells, inhibited cell proliferation and promoted apoptosis in a dose-dependent manner(P＜0.05). The highly-expressed miR-16-5p inhibited proliferation and promoted cell apoptosis(P＜0.05). Also, miR-16-5p targeted at PD-1 and negatively regulated PD-1 expression. Knockdown of PD-1 inhibited proliferation and promoted cell apoptosis(P＜0.05). PD-1 over-expression reversed the Anlotinib-mediated pro-proliferation and anti-apoptosis of miR-16-5p in A549 cells and H1299 cells(P＜0.05). Anlotinib significantly reduced tumor growth in vivo(P＜0.05). Conclusions Anlotinib may inhibit cell proliferation, anti-apoptosis, and reduce tumor growth for NSCLC, which is involved in miR-16-5p/PD-1 axis.

Key words: Anlotinib, non-small cell lung cancer, miR-16-5p, PD-1, cell proliferation