Notch1信号通路在人脑胶质瘤U87细胞增殖、凋亡中的作用及机制探讨

目的 观察Notch1信号通路在人脑胶质瘤U87细胞增殖和凋亡中的作用,并探讨其机制.方法 将U87细胞随机分为A、B、C组,分别加入Notch信号激活剂rhNF-κB、抑制剂DAPT及PBS共培养48 h.用MTT法检测细胞吸光度值(A值)观察细胞增殖情况,流式细胞术检测细胞凋亡率,RT-PCR、Western blot法分别检测U87细胞中的Notch1、Hes1、Bcl-2 mRNA及其蛋白.结果 培养24、48、72、96 h,A组细胞A值分别为0.185±0.007、0.398±0.012、0.735±0.015、1.083±0.031,B组分别为0.102±0.003、0.130±0.004、0.161±0.006、0.194±0.003,C组分别为0.167±0.005、0.265±0.008、0.496±0.011、0.737±0.016,A、B组与C组比较,P均＜0.05.A、B、C组细胞凋亡率分为0.96％±0.17％、26.51％±3.74％、8.76％±1.40％,A、B组与C组比较,P均＜0.05.与C组比较,A组细胞中Notch1、Hes1、Bcl-2 mRNA及其蛋白表达量均显著增加(P均＜0.05),B组细胞中Notch1、Hes1、Bcl-2 mRNA及其蛋白表达量均显著降低(P均＜0.05).结论 Notch1信号通路在人脑胶质瘤U87细胞增殖、凋亡中发挥了重要的调控作用,其机制可能与调节靶基因Hes1、抗凋亡蛋白Bcl-2表达有关.

Effect and mechanism of Notch1 signaling pathway on proliferation and apoptosis in human glioma U87 cells

Objective To investigate the effect of Notch1 signaling pathway on the proliferation and apoptosis of human glioma U87 cells and explore the mechanism.Methods Human human glioma U87 cells were randomly divided into 3 group.and cocultured with Notch activator rhNF-κB、Notch inhibitor DAPT and PBS.MTT assay was used to detect the effect on U87 cells proliferation,flow cytometry technique was also employed to detect the effects on apoptosis.RT-PCR and Western-blot were employed respectively to evaluate the mRNA and protein expressions of Notch1、Hes1 and Bcl-2 in U87 cells.Results A value of U87 cells cultured for 24,48,72,96 h were 0.185±0.007,0.398±0.012,0.735±0.015,1.083±0.031 in group A;0.102±0.003,0.130±0.004,0.161±0.006,0.194±0.003 in group B;0.167±0.005,0.265±0.008,0.496±0.011,0.737±0.016 in group C.A value in group A,B had significant difference copared with group C(all P<0.05).Cell apoptosis ratio in group A,B,C were 0.96%±0.17%,26.51%±3.74%,8.76%±1.40%.Cell apoptosis ratio in group A,B had significant difference copared with group C(all P<0.05).Notch1,Hes1,Bcl-2 mRNA and protein in group A,B had significant difference copared with group C(all P<0.05).Conclusions Notch1 sigaling pathway plays an important regulating role in the proliferation and apoptosis of human glioma U87 cells,and the probable mechanisms maybe that it can control the expressions of the target gene Hes1 and anti-apoptosis protein Bcl-2.