Abstract: Objective To study the effect and mechanism of verteporfin (VP), a Yes-associated protein (YAP) targeted inhibitor, combined with radiotherapy on the proliferation and migration behavior of gliomas. Methods After having been treated with radiotherapy, VP or VP combined with radiotherapy, glioma cell proliferation was detected by colony formation assay and CCK-8 assay. The molecular mechanism of VP combined with radiotherapy was explored using Western blotting. The cell migration was detected by wound healing assay. Results Cell proliferation decreased after radiotherapy or VP treatment alone. The inhibition effect of radiotherapy combined with VP treatment on cell proliferation was more significantly. After VP combined with radiotherapy treatment, the protein expression of cyclin-dependent kinase 4 (CDK4) and phospho-Rb (p-Rb) significantly decreased. Radiotherapy or VP treatment alone inhibited cell migration. The cell migration capacity decreased more significantly after radiotherapy combined with VP treatment. Conclusions VP can significantly improve the anti-tumor effect of radiotherapy on glioma. VP combined with radiotherapy can block cell cycle and cell proliferation by interfering the interaction of YAP - transcriptional enhanced associate domains (YAP-TEAD).