摘要
目的探讨土木香内酯对人胶质瘤U251细胞增殖、凋亡的影响及其作用机制。方法体外培养人胶质瘤U251细胞,取对数期生长的U251细胞分为空白对照组和土木香内酯(浓度分别为2.5、5.0、10.0μmol/L)组,每组各10个复孔。使用MTT法检测U251细胞的增殖抑制率,采用流式细胞术检测U251细胞的凋亡率,免疫印迹法检测PDK1/Akt/GSK3β蛋白表达,使用RTPCR检测c-Myc, Cyclin D1 mRNA的表达水平。结果随土木香内酯浓度增加,U251细胞增殖抑制率明显增高(P<0.05),细胞调亡率明显增高(P<0.05)。与空白对照组相比,木香内酯组U251细胞PDK1、Akt和GSK3β蛋白表达水平均无明显差异(P>0.05),但PDK1、Akt和GSK3β蛋白的磷酸化水平均显著降低(P<0.05)。随着土木香内酯浓度的增加,U251细胞c-Myc、CyclinD1mRNA的表达水平明显降低(P<0.05)。结论土木香内酯抑制人胶质瘤U251细胞的增殖并促进其凋亡,其作用机制可能是通过调节PDK1/Akt/GSK3β信号通路有关,且在一定范围内呈浓度依赖性。
Objective To explore the effect of alantolactone on the apoptosis of human glioma cell line U251 cells and its mechanism. Methods Human glioma U251 cells were cultured in vitro, and then they were divided into blank control group,experimental group I in which U251 cells were treated with 2.5 μmol/L alantolactone, experimental group Ⅱ in which U251 cells were treated with 5.0 μmol/L alanolastone and experimental group Ⅲ in which U251 cells were treated with 10.0 μmol/L alanolastone. The MTT assay was used to detect the proliferation of U251 cells and the apoptosis of U251 cells was detected by flow cytometry 24 hours after the treatment with different doses drug. The protein expressions of phophoinositid-dependent kinase(PDK1), protein kinase B(Akt)and glycogen synthase kinase-3β(GSK3β) in the U251 cells were detected by Western blot. The expressions of c-Myc and cyclin D1 mRNA were detected by RT-PCR. Results The results of MTT assay showed that the different doses of alantolactone effectively inhibited the proliferation of U251 cells(P<0.05), and the inhibition rate of U251 cells proliferation increased with the increase of the dose of alantolactone. Flow cytometry results showed that the apoptosis rates of the U251 cells were significantly higher in all the experimental groups than that in the blank control group(P<0.05), and the apoptosis of U251 cells induced by alantoctone is significantly dose-dependent(P<0.05). There were insignificant differences in the levels of PDK1, Akt and GSK3β protein expressions in U251 between the experimental group and blank control group and among all the experimental groups(P>0.05), but the levels of phosphorylated protein expression were significantly lower in all the experimental groups than that in the blank control group(P<0.05).The results of RT-PCR showed that the levels of c-Myc and cyclin D1 mRNA expressions in the U251 cells were significantly lower in the all the experimental groups than those in the blank control group(P<0.05)and the down-regulation of the levels of c-Myc and cyclin D1 induced by alantolactone was significantly dose-dependent(P<0.05). Conclusion It is suggested that alantolactone can inhibit the proliferation and promotes apoptosis of human glioma cell line U251 cells in a dose-dependent manner possibly by the regulation of the expressions of the proteins related to PDK1/Akt/GSK3β signaling pathway and cell cycle factors.
作者
张旭
田召辉
郭琳
娄永利
ZHANG Xu;TIAN Zhao-hui;GUO Lin;LOU Yong-li(Department of Neurosurgery,General Hospital,Pingmei Shenma Medical Group,Pingdingshan 467000 China;Department of Neurosurgery,Zhengzhou Municipal Central Hospital Affiliated to Zhengzhou University Zhengzhou,450000,China)
出处
《中国临床神经外科杂志》
2020年第1期32-35,共4页
Chinese Journal of Clinical Neurosurgery
基金
2016年河南省医学科技攻关计划(201602379)
