Glioma is one of the most destructive human tumours. Although standard treatment has improved the prognosis for glioma patients, the survival of glioma patients is still unsatisfactory. Avasimibe, an effective inhibitor of cholesterol acyltransferase 1 (ACAT1), has shown anti-tumour efficacy in many kinds of tumours. However, its role and related mechanism in glioma has not been fully elucidated. In the present study, we show that avasimibe effectively inhibits the proliferation, migration and invasion of glioma cell lines. Through LncRNA microarrays, we found that linc00339 levels were closely related to the anti-tumour effect of avasimibe. With the help of a series of functional assays, we show that avasimibe inhibits the proliferation, migration and invasion of glioma cell lines by suppressing linc00339 in vitro and in vivo. Our findings may provide a new approach for glioma therapy.