Department of Neurosurgery, The First Affiliated Hospital of Nanjing Medical University, China.
Acta Pharmacol Sin. 2012 Jul;33(7):935-40. doi: 10.1038/aps.2012.46. Epub 2012 Jun 11.
To examine the effects of anisomycin on glioma cells and the related mechanisms in vitro.
The U251 and U87 human glioblastoma cell lines were tested. The growth of the cells was analyzed using a CCK-8 cell viability assay. Apoptosis was detected using a flow cytometry assay. The expression of proteins and phosphorylated kinases was detected using Western blotting.
Treatment of U251 and U87 cells with anisomycin (0.01-8 μmol/L) inhibited the cell growth in time- and concentration-dependent manners (the IC(50) values at 48 h were 0.233±0.021 and 0.192±0.018 μmol/L, respectively). Anisomycin (4 μmol/L) caused 21.5%±2.2% and 25.3%±3.1% of apoptosis proportion, respectively, in U251 and U87 cells. In the two cell lines, anisomycin (4 μmol/L) activated p38 MAPK and JNK, and inactivated ERK1/2. However, neither the p38 MAPK inhibitor SB203580 (10 μmol/L) nor the JNK inhibitor SP600125 (10 μmol/L) prevented anisomycin-induced cell death. On the other hand, anisomycin (4 μmol/L) reduced the level of PP2A/C subunit (catalytic subunit) in a time-dependent manner in the two cell lines. Treatment of the two cell lines with the PP2A inhibitor okadaic acid (100 nmol/L) caused marked cell death.
Anisomycin induces glioma cell death via down-regulation of PP2A catalytic subunit. The regulation of PP2A/C exression by anisomycin provides a clue to further study on its role in glioma therapy.
在体外研究放线菌酮对神经胶质瘤细胞的作用及其相关机制。
采用 U251 和 U87 人神经胶质瘤细胞系进行实验。用 CCK-8 细胞活力检测法分析细胞生长情况。用流式细胞术检测细胞凋亡。用 Western blot 检测蛋白和磷酸化激酶的表达。
放线菌酮（0.01-8 μmol/L）处理 U251 和 U87 细胞，呈时间和浓度依赖性抑制细胞生长（48 h 的 IC50 值分别为 0.233±0.021 和 0.192±0.018 μmol/L）。放线菌酮（4 μmol/L）分别导致 U251 和 U87 细胞凋亡比例达到 21.5%±2.2%和 25.3%±3.1%。在两种细胞系中，放线菌酮（4 μmol/L）激活 p38 MAPK 和 JNK，抑制 ERK1/2。然而，p38 MAPK 抑制剂 SB203580（10 μmol/L）和 JNK 抑制剂 SP600125（10 μmol/L）均不能阻止放线菌酮诱导的细胞死亡。另一方面，放线菌酮（4 μmol/L）在两种细胞系中呈时间依赖性降低 PP2A/C 亚基（催化亚基）的水平。用蛋白磷酸酶 2A（PP2A）抑制剂冈田酸（100 nmol/L）处理两种细胞系，引起明显的细胞死亡。
放线菌酮通过下调 PP2A 催化亚基诱导神经胶质瘤细胞死亡。放线菌酮对 PP2A/C 表达的调控为进一步研究其在神经胶质瘤治疗中的作用提供了线索。
