新型SMO抑制剂SI-4650对人神经胶质瘤U87MG细胞增殖、凋亡和自噬能力的影响

目的研究新型精胺氧化酶(spermine oxidase, SMO)小分子抑制剂SI-4650对人胶质瘤U87MG细胞增殖、凋亡和自噬的影响及其分子机制。方法 MTT法检测细胞增殖情况;化学发光法检测SMO和乙酰多胺氧化酶(N^1-acetylpolyamine oxidase, APAO)的酶活性;高效液相色谱法(HPLC)检测细胞内多胺含量;Transwell法分析U87MG细胞的迁移能力;PI单染后结合流式细胞术分析细胞周期;PI/FITC-Annexin V双染后结合流式细胞术、Western blot分析细胞凋亡;激光共聚焦显微镜(LSCM)和Western blot分析细胞自噬。结果 SI-4650可明显抑制U87MG细胞内SMO和APAO的酶活性,干扰多胺代谢,并降低U87MG细胞内总多胺含量。用SI-4650处理能抑制U87MG细胞的增殖和迁移能力,使细胞发生G_0/G_1周期阻滞,并诱导U87MG细胞发生凋亡和自噬性死亡。结论 SI-4650具有杀伤神经胶质瘤U87MG细胞的药理活性,其机制可能与干扰多胺代谢和诱导细胞凋亡和自噬相关。

Effects of novel SMO inhibitor SI-4650 on proliferation,apoptosis and autophagy of human glioma U87MG cells

Aim To investigate the effects of a novel spermine oxidase(SMO)inhibitor SI-4650on proliferation,apoptosis and autophagy of human glioma U87MG cells as well as its possible mechanism.Methods MTT assay was used to detect cell proliferation.The chemiluminescence assay was used to determine enzyme activities of SMO and N^1-acetylpolyamine oxidase(APAO).High performance liquid chromatography(HPLC)was performed to detect cellular polyamine content.Transwell assay was used to evaluate the migration ability of U87MG cells.PI single-staining/flow cytometry(FCM)were used to analyze cell cycle.PI/FITC-Annexin V double-staining/FCM and Western blot were used to analyze apoptosis.Confocal laser scanning microscopy(LSCM)and Western blot were used to analyze autophagy.Results SI-4650could significantly inhibit the activities of SMO and APAO,interfere with polyamine metabolism and reduce the content of total polyamine in U87MG cells.Treatment of U87MG cells with SI-4650inhibited the proliferation and migration,caused G0/G1cell cycle arrest and induced apoptotic and autophagic cell death.Conclusions SI-4650has the pharmacological activity of killing glioma U87MG cells,and its mechanism may be related to the interference of polyamine metabolism and induction of cell apoptosis and autophagy.