UM-164 Inhibits Proliferation Of Human Glioma Cells By Reducing The Activity Of YAP

Objective : As the most common primary craniocerebral malignant tumor in clinic,glioma has the characteristics of high incidence,high mortality and high recurrence rate.The current routine treatment scheme of glioma is the most safe degree of surgical resection and synchronous radiotherapy and chemotherapy,but the extremely high invasion characteristics and resistance to radiotherapy and chemotherapy make the routine treatment of glioma not significant,and the survival time of the patients is still short.Therefore,targeted therapy for pathogenic genes and related signaling pathways has become one of the important methods for the treatment of glioma.Src is the first proto-oncogene found in humans,and its encoded protein is a non-receptor tyrosine kinase that plays an important role in promoting the proliferation,invasion and migration of glioma cells.Researches have confirmed that interfering with the expression of Src can inhibit the proliferation of glioma cells.UM-164 is a newly developed and efficient Src inhibitor,which has higher binding force with Src than traditional inhibitors.Nevertheless,there are no relevant reports UM-164 affect the proliferation of glioma cells.To investigate the effect of UM164,a Src inhibitor,on the proliferation of human glioma cells,and to elucidate the related mechanism,so as to lay a theoretical foundation for the development of new drugs for glioma targeted therapy.Methods : Human glioma cells SF767 were divided into blank control group and UM-164-treated group.Cell Counting kit-8(CCK-8)was employed to detect the viability of SF767 cells.Colony forming ability of SF767 cells was detected by colony formation assay.The effect of UM-164 on SF767 cells proliferation was analyzed by BrdU and Ki67 incorporation assays.Cell cycle distribution and apoptosis were analyzed by flow cytometry(FCM),and then expression of Src,p38,YAP and Cyclin D1 were detected by Western blotting.The localization of YAP was observed by immunofluorescence assay.Results : UM-164 could significantly inhibited SF767 cell viability in a time-and dose-dependent manner(P<0.05),and UM-164 could inhibit colony formation of SF767cells(P<0.01).BrdU and Ki67 incorporation assays showed the numbers of BrdU and Ki67 positive cells marked with BrdU or Ki67 were significantly reduced after the treatment with UM-164(P<0.01).FCM results indicated that UM-164 could induce SF767 cell arrest in phase G0/G1(P<0.01),but could not induce apoptosis of SF767 cells.The Western blotting showed that the expression of p-Src,p-p38,Cyclin D1 and p-YAP(S397)were significantly decreased with UM-164 treatment while the expression of Cyclin D1 was significantly increased(P<0.01).Immunofluorescence results showed that UM-164 induced YAP to exclude from the nucleus.Conclusion : UM-164 can inhibit the proliferation of human glioma cells,and the mechanism may be related to the reduction of YAP activity by UM-164.