司美替尼对恶性周围神经鞘瘤细胞增殖和凋亡的影响

The effect of selumetinib on cell proliferation and apoptosis of malignant peripheral nerve sheath tumor

目的:探讨MEK1/2抑制剂司美替尼(selumetinib)对恶性周围神经鞘瘤(MPNST)细胞增殖和凋亡的影响及其机制。方法:用不同浓度的司美替尼处理细胞株ST88-14和STS26T,通过CCK-8、克隆形成、Transwell、细胞划痕、流式细胞术检测司美替尼对细胞增殖、凋亡、迁移和侵袭的影响。实时荧光定量PCR检测多梳抑制复合物2(PRC2)核心组分的mRNA表达水平。免疫印迹试验检测组蛋白3上的第27位赖氨酸的三甲基化(H3K27me3)的表达。结果:司美替尼抑制ST88-14、STS26T细胞增殖(t=16.44,P<0.05;t=16.21,P<0.05)、侵袭(t=10.51,P<0.05;t=8.44,P<0.05)、迁移(t=4.02,P<0.05;t=2.21,P<0.05);与对照组相比,10μmol/L和20μmol/L司美替尼可使ST88-14、STS26T细胞凋亡率显著升高(t=14.64、10.10,均P<0.05;t=3.06、13.10,均P<0.05);与对照组相比,司美替尼作用于ST88-14、STS26T细胞48 h,PRC2核心组分SUZ12和EZH2的mRNA水平显著升高(t=13.39、16.84,均P<0.05;t=6.10、12.93,均P<0.05);与对照组相比,20μmol/L司美替尼可显著增加ST88-14,STS26T细胞H3K27me3的表达(t=12.82,P<0.05;t=18.78,P<0.05)。结论:司美替尼通过促进H3K27me3表达抑制MPNST细胞的增殖,促进细胞凋亡。

Objective:To investigate the effects of selumetinib,a MEK1/2 inhibitor,on cell proliferation and apoptosis of malignant peripheral nerve sheath tumor(MPNST)and its mechanism.Methods:Cell lines ST88-14 and STS26T were treated with selumetinib at different concentrations,and the effects of selumetinib on cell proliferation,apoptosis,migration and invasion were detected by CCK-8 assay,colony formation assay,Transwell assay,cell scratch assay and flow cytometry.The mRNA expression level of each core component of PRC2 was detected by real-time fluorescence quantitative PCR.Western blotting assay was used to detect the expression of trimethylation of lysine 27 on histone 3(H3K27me3).Results:Selumetinib inhibited the proliferation(t=16.44,P<0.05;t=16.21,P<0.05),invasion(t=10.51,P<0.05;t=8.44,P<0.05)and migration(t=4.02,P<0.05;t=2.21,P<0.05)of ST88-14,STS26T cells.Compared with the control group,the apoptosis rates of ST88-14 and STS26T cells were significantly increased when drug concentrations were 10μmol/L and 20μmol/L(t=14.64,10.10,both P<0.05;t=3.06,13.10,both P<0.05).Compared with the control group,the mRNA levels of SUZ12 and EZH2,the core components of PRC2,were significantly increased by selumetinib for 48 h after treatment with ST88-14 and STS26T cells(t=13.39,16.84,both P<0.05;t=6.10,12.93,both P<0.05).Compared with the control group,20μmol/L selumetinib significantly increased the expression of H3K27me3 in ST88-14 and STS26T cells(t=12.82,P<0.05;t=18.78,P<0.05).Conclusion:Selumetinib can inhibit the proliferation of MPNST cell lines and promote apoptosis by promoting the expression of H3K27me3.