Anlotinib inhibits glioma cell proliferation, migration, and invasion by suppressing c-MET/AKT/mTOR signaling and altering autophagy-associated protein expression.

Abstract

Glioma remains difficult to treat because malignant cells show rapid proliferation, diffuse invasion, and frequent recurrence. This study investigated the direct antitumor effects of anlotinib in human glioma cells and examined whether these effects were associated with hepatocyte growth factor receptor/protein kinase B/mechanistic target of rapamycin (c-MET/AKT/mTOR) signaling, apoptosis-associated responses, mesenchymal-transition-related proteins, and autophagy-associated proteins. U87 and U251 cells were treated with anlotinib in vitro. Cell viability was assessed using Cell Counting Kit-8 assays, migration and invasion were evaluated using wound-healing and Transwell assays, Hoechst 33342 staining was used for qualitative assessment of apoptosis-associated nuclear morphology; and protein expression was analyzed by western blotting. Anlotinib reduced cell viability in a concentration-dependent manner, with estimated 24 h IC50 values of 68.00 μM for U87 cells and 45.53 μM for U251 cells. At lower concentrations, anlotinib reduced wound closure and Transwell invasion, decreased Vimentin, N-cadherin, and β-catenin expression, increased B-cell lymphoma 2 (BCL-2)-associated X protein, decreased BCL-2, altered poly(ADP-ribose) polymerase/caspase-9 expression, reduced total c-MET expression, and decreased AKT/mTOR phosphorylation. Analysis of autophagy-associated proteins showed an increased LC3-II/LC3-I and time-dependent p62 changes. Anlotinib may suppress malignant glioma-cell phenotypes in vitro. These phenotypic changes were associated with alterations in c-MET/AKT/mTOR pathway proteins, apoptosis-associated responses, mesenchymal-transition-related proteins, and autophagy-associated protein expression.