小分子化合物WAY-312通过周期阻滞与凋亡诱导抑制胶质母细胞瘤干细胞增殖及自我更新OA

Objective:This study investigated the inhibitory activity and potential mechanism of WAY-312,a small-molecule compound derived through screening of a small-molecule compound library,against the growth and self-renewal capacity of glioblastoma stem cells(GSCs).Methods:Systematic compound screening was conducted on the small molecule compound library through tumor sphere formation assays and cell viability assays;the small molecule inhibitor WAY-302386(named WAY-312 according to its compound number)obtained through screening was used as the research object.CCK-8 method,Western blot,flow cytometry,and Caspase-Glo 3/7 activity detection method were used to detect the effects of WAY-312 on the proliferation ability,stemness marker expression,cell cycle distribution,and apoptosis level of glioblastoma stem cells at different concentrations.The regulatory effect of WAY-312 on GSC stemness maintenance and self-renewal ability was clarified,and the mechanism of WAY-312 on GSC cell cycle progression and apoptosis was further elucidated.Results:WAY-312 can effectively inhibit the survival of GSCs(P<0.05);WAY-312 significantly inhibits the number and size of tumor spheroids formed in GSCs(P<0.05),and treatment with WAY-312 reduces the expression levels of stemness marker proteins SOX2,OLIG2,and BMI1;WAY-312 can effectively inhibit the self-renewal ability of GSCs(P<0.05);WAY-312 induces GSC cell cycle arrest at the G2/M phase by regulating the expression of cell cycle-related proteins Cyclin B1,CDK4,and CDK6(P<0.05);WAY-312 induces apoptosis in GSCs,increases the expression of apoptosis-related proteins cleaved-PARP and BAX,reduces the expression of BCL2,and enhances Caspase-3/7 activity in GSCs after treatment with WAY-312.Conclusion:WAY-312 can inhibit GSC proliferation,stemness maintenance,and self-renewal ability,induce GSC cell cycle arrest at G2/M phase,and promote cell apoptosis in a concentration-dependent manner.