多胺类似物DENSPM对人胶质母细胞瘤SNB19细胞生长的影响

Influence of polyamine analogue DENSPM on cell growth of human SNB19 glioblastoma cells

目的探讨多胺类似物DENSPM对人胶质母细胞瘤sNB19细胞生长的影响。方法采用MTS法、流式细胞术、高效液相色谱法和逆转录-聚合酶链反应,分别检测经DENSPM处理后的SNB19细胞存活率、细胞周期变化和细胞内过氧化氢水平,以及细胞内多胺水平和鸟氨酸脱羧酶、亚精胺/精胺-N1-乙酰基转移酶、多胺氧化酶mRNA表达变化。结果经DENSPM处理后,SNB19细胞存活率随着药物浓度的增加逐渐降低(F=81.915,P=0.001),并于体外培养72h在细胞增殖周期中出现典型的亚凋亡峰;细胞内腐胺、亚精胺和精胺水平显著下降,而乙酰亚精胺和乙酰精胺水平略有上升(均P<0.01);鸟氨酸脱羧酶表达水平下降(P<0.01),而亚精胺/精胺-N1-乙酰基转移酶和多胺氧化酶水平上升(P<0.05或P<0.01);但细胞内过氧化氢水平与DENSPM处理前差异无统计学意义(P>0.05)。结论 DENSPM可抑制人胶质母细胞瘤SNB19细胞的生长并诱导其发生凋亡,而诱导细胞凋亡的可能机制是肿瘤细胞内多胺表达水平下降。DENSPM可能具有治疗胶质母细胞瘤的潜在临床应用价值。

Objective To investigate the influence of polyamine analogue DENSPM （N1, Nlldiethylnorspermine） on cell growth of human SNB19 glioblastoma cells. Methods DENSPM was added into the culture medium of SNB19 cells. MTS assay was used to assess cell survival rate. The change of the cell cycle and the value of intracellular H202 were evaluated by flow eytometry. The level of polyamine was measured by HPLC. The expression of ornithine deearboxylase （ODC）, spermidine/spermine N（1）- aeetyltransferase （SSAT）, polyamine oxidasc （PAO） mRNA was identified by quantitative RT- PCR. Results Cell survival rate of SNB19 cells decreased markedly after DENSPM treatment （F = 81.915, P = 0.001）. DENSPM treatment led to tbe appearance of typical sub-G peak in flow eytometry assay. The levels of putrescine, spermidine and spermine reduced （P 〈 0.01）, while the level of acetylspermidine and acetylspermine slightly increased （P 〈 0.01）. Meanwhile, the mRNA expression of ODC decreased （P 〈 0.01）. The level of SSAT and PAO increased （P 〈 0.01, P 〈 0.05, respectively）. However, the value of intracellular H202 was unchanged （P 〉 0.05）. Conclusion The polyamine analogue DENSPM inhibits cell growth and induces apoptosis of SNB19 glioblastoma cells. Reduction of polyamine in SNB19 cells is the possible meehansm of apoptosis. DENSPM may be a potential medication for glioblastoma.