多胺类似物DENSPM对人胶质瘤LN229细胞增殖的抑制作用

Plyamine analogue DENSPM inhibits cell proliferation of human LN229 glioblastoma cells

目的探讨多胺类似物二乙基去甲精胺(DENSPM)对人的胶质母细胞瘤LN229细胞的抑制作用。方法用DENSPM处理LN229细胞后,采用单溶液细胞增殖检测(MTS)法、细胞流式术、高效液相色谱法、定量逆转录酶-聚合酶链反应(qRT-PCR),分别检测其细胞增殖率、细胞周期变化和细胞内过氧化氢水平、细胞内多胺表达水平、亚精胺/精胺N1-乙酰基转移酶(SSAT)、多胺氧化酶(PAO)、鸟氨酸脱羧酶(ODC)mRNA表达变化。结果 LN229细胞经DENSPM处理后,其存活率随着药物浓度的增加而逐渐降低(F=124.051,P<0.001),并于体外培养72 h后在细胞增殖周期中出现典型的亚凋亡峰;细胞内过氧化氢水平于72 h后显著升高(F=57.27,P<0.001);细胞内腐胺、亚精胺和精胺水平显著下降(P≤0.001),而乙酰亚精胺和乙酰精胺水平显著上升(P<0.001);亚精胺/精胺-N1-乙酰基转移酶、多胺氧化酶和鸟氨酸脱羧酶水平上升(P<0.001)。结论 DENSPM可能通过降低肿瘤细胞内多胺表达水平,来抑制LN229细胞的生长并诱导其发生凋亡。

Objective To explore the inhibition of polyamine analogue N1, N11-diethylnorspermine （DENSPM） on cell growth of human LN229 glioblastotm cells. Methods LN229 cells were cultured in medium with dialyzed serum. After treatment with DENSPM, one solution cell proliferation assay （MTS） was used to assess cell viability. The change of cell cycle and the expression levels of intracellular H2O2 were assessed by flow cytometry. The levels of polyamine were measured by high performance liquid chromatography （HPLC）. The expression levels of spermidine/spennine N （ 1 ）-acetyltransferase （SSAT）, polyamine oxidase （PAO）, and omithine decarboxylase （ODC） mRNA were detected by quantitative reverse transcription-polymerase chain reaction （qRT-PCR）. Results Cell survival rate of LN2~ cells was decreased rmrkedly with the increase of DENSPM concentration （F = 124. （151, P 〈0.001）. The typical sub G1 peak appeared after cultured in vitro for 72 h in flow cytometry assay. Compared with the control groups, the levels of I-I202 were increased markedly. The levels of putrescine, spermidine and spermine declined markedly （P≤0.001）, while the levels of acetylspermidine and acetylspermine were increased （P 〈0.1301 ）. Meanwhile, the mRNA expressions of SSAT, PAO and ODC were increased （ P 〈 0. 001 ）. Condusion The polyamine analogue DENSPM can reduce the intracellular polyamine levels and thus inhibits the cell growth and induces the apoptosis of LN229 glioblastoma cells.