In this study, the function of long non-coding RNA SOX21 antisense RNA 1 (SOX21-AS1) in the progress of glioma was explored. RNA and protein levels were measured via quantitative reverse transcription-PCR (qRT-PCR) and western blot analysis. In addition, we examined cell proliferation, apoptosis, migration and invasion. The interaction between SOX21-AS1 (PAK7) and miR-144-3p was determined via RNA immunoprecipitation (RIP) assay and Luciferase reporter assay. SOX21-AS1 was upregulated in glioma tissues and cells. SOX21-AS1 knockdown was carried out in glioma cells (U251 and U87 cells). Moreover, in vitro, SOX21-AS1 knockdown repressed proliferation, migration, invasion and enhanced apoptosis in glioma cells. In vivo, SOX21-AS1 knockdown suppressed tumor growth in mice. In addition, SOX21-AS1 could sponge miR-144-3p, which was determined to bind to PAK7. miR-144-3p knockdown promoted proliferation, migration, invasion and inhibited cell apoptosis. Importantly, the effects of SOX21-AS1 knockdown-induced proliferation, migration, invasion, and apoptosis were alleviated in glioma cells co-transfected with SOX21-AS1 and miR-144-3p knockdown. Furthermore, miR-144-3p knockdown also attenuated Wnt/β-catenin pathway-associated protein levels induced by SOX21-AS1 knockdown. These results indicated that SOX21-AS1/miR-144-3p/PAK7 axis played an oncogenic role in glioma cells by regulating Wnt/β-catenin pathway, which suggests a rational therapeutic strategy for glioma.