Effects of NPS-2143 on proliferation and migration ability of glioma cells and its mechanism
Objective To explore the effect and mechanism of NPS-2143 on proliferation and migration of glioma U87 and U251,and Hs683 cells.Methods The glioma cells were divided into 3 groups:the Control group(without medical treatment),the 6 μmol/L and the 8 μmol/L NPS-2143 treatment group.The effects of NPS-2143 on the viability and proliferation of U87 and U251 cells were investigated by using cell counting kit-8(CCK-8)and colony formation experiments after the treatment with different concentrations of NPS-2143.Flow cytometer(FCM)was used to detect the effects of different concentrations of NPS-2143 on cycle and apoptosis of U87 and U251 cells.The expression levels of apoptosis-related proteins(Bax,Caspase 3)and cell cycle related proteins(P21,Cyclin D1)in U87 and U251 were detected by Western blot;The migration capability of U87 and Hs683 cells treated with different concentrations of NPS-2143 was detected by wound healing and Transwell experiments.The expression levels of epithelial-mesenchymal transition(EMT)related proteins(N-cadherin,MMP2,MMP9)in U87 and Hs683 cells were detected by Western blot;the expression levels of autophagy proteins(LC3,P62)and AKT-mTOR pathway related proteins(p-AKT,p-mTOR)in U87 and U251 cells were detected by Western blot.The changes of the number of autophagy vesicles in glioma U87 cells treated with NTP-2143 were observed by transmission electron microscopy.Results The viability and proliferation ability of glioma cells were reduced,with dependence of time and concentration(P＜0.01),the proportaion of G1 phase and apoptotic glioma cells increased(P＜0.05)and the expression level of P21,Bax,and Caspase 3 increased(P＜0.01)while the level of Cyclin D1 decreased(P＜0.01)in the NPS-2143 treatment group,compared with the Control group.NPS-2143 inhibited the migration ability of glioma cells(P＜0.01)and decreased the expression levels of N-cadherin,MMP2,and MMP9(P＜0.01),and also increased the expression levels of autophagy proteins LC3,P62,p-AKT,and p-mTOR(P＜0.01),compared with the Control group.The number of autophagy vesicles obviously increased in glioma U87 cells treated with NPS-2143,compared with the Control group(P＜0.01).Conclusion NPS-2143 may inhibit the proliferation and migration of glioma cells by blocking autophagy through mediating the AKT-mTOR pathway.
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