UNBS5162对脑胶质瘤细胞增殖和转移的抑制作用及机制

Effect of UNBS5162 on proliferation and metastasis of glioblastoma cells and its mechanism

目的通过体外实验,研究UNBS5162对脑胶质瘤细胞增殖和转移的抑制作用,并对其分子机制进行初步探究。方法采用10μmol/L UNBS5162处理脑胶质瘤细胞U251细胞为实验组,二甲基亚砜处理为对照组,分别应用CCK8实验、Transwell实验、流式细胞凋亡实验检测UNBS5162对U251细胞增殖、迁移和侵袭、凋亡的影响,应用蛋白免疫印迹法检测PI3K/AKT信号通路相关蛋白的表达水平。结果实验组细胞增殖能力低于对照组(P<0.05);并且其细胞迁移数和侵袭数均低于对照组[(18±5)个比(32±2)个,P<0.05;(30±2)个比(78±4)个,P<0.05];与对照组相比,实验组细胞凋亡率提高[(24.46±2.03)%比(5.75±0.74)%,P<0.05],且抗凋亡蛋白Bcl-2表达量降低、促凋亡蛋白Bax和Caspase-3表达量增加(P<0.05)。与对照组相比,PI3K/AKT信号通路的关键蛋白AKT和m TOR的磷酸化水平受到抑制,其下游p70S6K蛋白表达水平也相应降低(P<0.05)。结论 UNBS5162通过促进细胞凋亡对脑胶质瘤细胞的增殖和转移具有抑制作用,其机制与UNBS5162可以抑制PI3K/AKT信号通路的激活有一定相关性。

Objective To investigate the effect of UNBS5162 on glioblastoma cell proliferation and metastasis in vitro,and to preliminarily explore its molecular mechanism. Methods Glioblastoma U251 cells were treated with 10 μmol/L UNBS5162（ as experimental group） or dimethyl sulfoxide（ as control group）,respectively. After treatment,the effect of UNBS5162 on cell proliferation,migration,invasion,and apoptosis of U251 cells were examined by CCK8 assay,Transwell assay and flow cytometry assay,respectively. The protein level of PI3K/AKT pathway-related proteins was examined by Western blot. Results The proliferation ability of UNBS5162 treated cells was significantly lower than that of control group（ P〈0. 05）. Compared with control group,the number of migrated cells [（ 18 ± 5） vs（ 32 ± 2）,P〈0. 05] and invaded cells [（ 30 ± 2） vs（ 78 ± 4）,P〈0. 05] all significantly decreased in UNBS5162 treated group. Moreover,the apoptotic rate of UNBS5162 treated group was significantly promoted compared with that of control group [（ 24. 46 ± 2. 03） % vs（ 5. 75 ± 0. 74） %,P〈0. 05],and compared with control group,the expression level of anti-apoptotic protein Bcl-2 was down-regulated,while the expression levels of pro-apoptotic protein Bax and Active Caspase-3 were markedly up-regulated. Furthermore,compared with control group,the phosphorylation levels of the key proteins in PI3K/AKT pathway,AKT and mTOR,were all inhibited,and the downstream p70S6K protein expression also decreased. Conclusion UNBS5162 has significant inhibitory effect on cell proliferation and metastasis of glioblastoma cells by promoting apoptosis,and its mechanism might be related to the activity inhibition of PI3K/AKT signaling pathway in the glioblastoma cells.