| Objective To explore the effects of selective monoamine oxidase B(MAO-B) inhibitor TM-5 on autophagy and apoptosis of glioma U87 cells. Methods The normally cultured glioma cells U87 were randomly divided into control group(no treatment), 5, 10, 25, 50and 100 μmoL/L TM-5 groups(5, 10, 25, 50 and 100 μmoL/L of TM-5), with 3 experimental pores in each group. The cells activity was detected by MTT. Apoptosis was detected by flow cytometry. The expressions of pro-apoptosis protein Bax and anti-apoptosis protein Bcl-2genes were detected by real-time fluorescent quantitative polymerase chain reaction(qRT-PCR). The expressions of apoptosis-relatedproteins(Bax, cleaved caspase-3) and autophagy related markers(Beclin-1, LC3-II, P62) were detected by Western blot(WB). Results Compared with the control group, the survival rate of U87 cells decreased in 10, 25, 50 and 100 μmoL/L TM-5 groups(P<0.05). The follow-up experiments were performed under certain concentration(5, 10 and 25 μmoL/L, cells survival rate >50%). Compared with the control group, relative apoptosis rate of U87 cells and relative expression levels of LC3-Ⅱand Bax proteins increased in 10 and 25 μmoL/L TM-5 groups(P<0.05).Compared with control group, relative expression levels of pro-apoptosis gene Bax mRNA, Beclin-1 and cleaved caspase-3 proteins increased, while relative expression levels of anti-apoptosis gene Bcl-2mRNA and P62 protein decreased in 5, 10 and 25 μmoL/L TM-5 groups(P<0.05).Conclusion TM-5 can inhibit the survival of glioma U87 cells, up-regulate Beclin-1, LC3-Ⅱ, Bax and cleaved caspase-3, down-regulate Bcl-2 and P62, and induce autophagy and apoptosis. |
