Glioma is one of the most common tumors in China and chemotherapy is critical for its treatment. Recentstudies showed that benzyl isothiocyanate (BITC) could inhibit the growth of glioma cells, but the mechanisms arenot fully understood. This study explored the inhibitory effect of BITC on invasion and angiogenesis of U87MGhuman glioma cells in vitro and in vivo, as well as potential mechanisms. It was found that BITC could inhibitinvasion and angiogenesis of human glioma U87MG cells by inducing cell cycle arrest at phase G2/M. It alsowas demonstrated that BITC decreased expression of cyclin B1, p21, MMP-2/9, VE-cadherin, CD44, CXCR4and MTH1, the activity of the telomerase and PKCζ pathway. Microarray analysis was thus useful to explorethe potential target genes related to tumorigenic processes. BITC may play important roles in the inhibition ofinvasion and angiogenesis of human glioma cells.