纳洛酮对神经胶质瘤U87细胞的抗肿瘤作用及机制研究

Antitumor Effect and Mechanism of Naloxone on Glioma U87 Cells

目的探讨纳洛酮(Naloxone,NA)对神经胶质瘤U87细胞的抗肿瘤作用及其可能的机制。方法培养U87细胞,将U87细胞分为对照组和实验组(U87+NA),采用CCK-8细胞增殖实验检测NA对U87细胞活性的影响并确定最佳用药浓度;运用Western blot检测U87细胞中热休克蛋白60(HSP60)、热休克转录因子1(HSF-1)、半胱氨酸-天冬氨酸蛋白酶3(Caspase-3)、核因子-κB(NF-κB)和Toll样受体4(TLR-4)的表达水平;通过免疫荧光技术进一步检测U87细胞在加入NA前、后U87细胞生长状况的变化。结果与对照组相比,0.5～4.0μM的NA处理U87细胞后,神经胶质瘤U87细胞的活性均降低(P<0.05)。实验组NF-κB和TLR-4的表达较对照组减少,HSP60、Caspase-3的表达增加(P均<0.05)。与对照组相比,0.5μM NA作用U87细胞48h后抑制了U87细胞的生长(P<0.05)。结论纳洛酮可促进U87细胞的凋亡,可能与HSP60高表达以及NF-κB、TLR-4低表达有关。

Objective To investigate the antitumor effect of naloxone(NA)on glioma U87 cells and its mechanism.Methods U87 glioma cells were cultured,and divided into normal control group and experimental group(U87+NA).CCK-8 cell proliferation assay was used to detect the influence of NA on the activity of U87 cells and determine the optimal concentration.The expression levels of heat shock protein 60(HSP60),heat shock transcription facto1(HSF-1),cysteine-aspartate protease 3(caspase-3),nuclear factor-κB and toll-like receptor 4(TFR-4)in U87 cells were detected by Western blot.Further changes in the growth of U87 cells before and after NA treatment were measured by immunofluorescence technique.Results Compared with the control group,the activity of U87 cells treated with 0.5-4.0μM NA decreased(P<0.05).The expression of NF-κB and TLR-4 was decreased and the expression of HSP60 and caspase-3 was increased(P<0.05).Compared with the control group,0.5μM NA inhibited the growth of U87 cells after 48h(P<0.05).Conclusion Naloxone may promote the apoptosis of glioma U87 cells by increasing the expression of HSP60 and Caspase-3 and decreasing the expression of NF-κB and TLR-4.