Survivin selective inhibitor YM155 induce apoptosis in SK-NEP-1 Wilms tumor cells.
Survivin selective inhibitor YM155 induce apoptosis in SK-NEP-1 Wilms tumor cells.
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生存素选择性抑制剂 YM155 诱导 SK-NEP-1 Wilms 肿瘤细胞凋亡
DOI：
10.1186/1471-2407-12-619
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发表时间：
2012-12-26
期刊：
影响因子：
3.8
通讯作者：
Pan J
中科院分区：
文献类型：
--
作者：
Tao YF;Lu J;Du XJ;Sun LC;Zhao X;Peng L;Cao L;Xiao PF;Pang L;Wu D;Wang N;Feng X;Li YH;Ni J;Wang J;Pan J
Survivin是凋亡抑制蛋白家族的一员，是有丝分裂和细胞程序性死亡的关键调节因子。YM155是一种新型的分子靶向剂，它抑制Survivin的表达，Survivin在许多肿瘤类型中过表达。本研究的目的是检测YM155对SK-NEP-1细胞的抗肿瘤活性。方法体外培养SK-NEP-1细胞，用四甲基偶氮唑盐比色法和裸鼠实验检测YM155的抗肿瘤活性。用膜联蛋白V/碘化丙啶染色和流式细胞仪检测细胞培养中的细胞凋亡。用实时荧光定量聚合酶链式反应芯片分析YM155处理的肿瘤细胞的基因表达谱。然后使用MEV(多实验视图)集群软件对表达数据进行分析。结果YM155处理SK-NEP-1细胞后，对SK-NEP-1细胞增殖的抑制作用呈剂量依赖性。Annexin V分析、细胞周期和caspase-3活性检测表明，YM155可诱导SK-NEP-1细胞发生凋亡。YM155显著抑制SK-NEP-1移植瘤的生长(YM155 5 mg/kg：1.45±0.77 cm~3；YM155 10 mg/kg：0.95±0.55 cm~3)，与DMSO组(3.70±2.4 cm~3)和PBS组(3.78±2.20 cm~3，ANOVA<；0.01)相比。YM155治疗组肿瘤重量(YM155 5 mg/kg：1.05±0.24 g；YM155 10 mg/kg：0.72±0.17 g)与DMSO组(2.06±0.38 g)或PBS组(2.36±0.43 g，ANOVA P<；0.01)比较，差异有统计学意义(P<0.01)。实时定量聚合酶链式反应芯片分析显示，YM155治疗后，试验组与对照组之间有32个基因显著上调，54个基因显着下调。独创性路径分析(IPA)显示，细胞死亡是评分最高的网络，有65个焦点分子，显着性得分为44。IPA分析还将差异表达的基因归类为与细胞死亡、细胞功能维持、细胞形态、碳水化合物代谢以及细胞生长和增殖相关的生物机制。死亡受体信号转导通路(3.87E-19)、TNFR1信号转导通路、HIV1诱导细胞凋亡信号转导通路、细胞凋亡信号转导通路以及肿瘤发生的分子机制是最重要的四条途径。IPA分析还显示上调的顶级分子是BBC3、BIRC3、BIRC8、BNIP1、CASP7、CASP9、CD5、CDKN1A、CEBPG和Col4A3，下调的顶级分子是ZNF443、UTP11L、TP73、TNFSF10、TNFRSF1B、TNFRSF25、TIAF1、STK17A、SST和SPP1，上游调控因子是NR3C1、TP53、地塞米松、肿瘤坏死因子和Ak.YM155治疗SK-NEP-1移植瘤疗效显著，毒副作用小。实时定量聚合酶链式反应芯片分析首次显示了YM155处理后染料调控基因的表达谱。IPA分析也揭示了YM155治疗的新分子机制，如NR3C1和地塞米松可能是YM155治疗的新靶点。我们的结果可能为研究YM155诱导细胞凋亡的分子机制提供新的线索。
BackgroundSurvivin, a member of the family of inhibitor of apoptosis proteins, functions as a key regulator of mitosis and programmed cell death. YM155, a novel molecular targeted agent, suppresses survivin, which is overexpressed in many tumor types. The aim of this study was to determine the antitumor activity of YM155 in SK-NEP-1 cells.MethodsSK-NEP-1 cell growthin vitroandin vivowas assessed by MTT and nude mice experiments. Annexin V/propidium iodide staining followed by flow cytometric analysis was used to detect apoptosis in cell culture. Then gene expression profile of tumor cells treated with YM155 was analyzed with real-time PCR arrays. We then analyzed the expression data with MEV (Multi Experiment View) cluster software. Datasets representing genes with altered expression profile derived from cluster analyses were imported into the Ingenuity Pathway Analysis tool.ResultsYM155 treatment resulted in inhibition of cell proliferation of SK-NEP-1cells in a dose-dependent manner. Annexin V assay, cell cycle, and activation of caspase-3 demonstrates that YM155 induced apoptosis in SK-NEP-1 cells. YM155 significantly inhibited growth of SK-NEP-1 xenografts (YM155 5 mg/kg: 1.45 ± 0.77 cm3; YM155 10 mg/kg: 0.95 ± 0.55 cm3) compared to DMSO group (DMSO: 3.70 ± 2.4 cm3) or PBS group cells (PBS: 3.78 ± 2.20 cm3, ANOVA P < 0.01). YM155 treatment decreased weight of tumors (YM155 5 mg/kg: 1.05 ± 0.24 g; YM155 10 mg/kg: 0.72 ± 0.17 g) compared to DMSO group (DMSO: 2.06 ± 0.38 g) or PBS group cells (PBS: 2.36 ± 0.43 g, ANOVA P < 0.01). Real-time PCR array analysis showed between Test group and control group there are 32 genes significantly up-regulated and 54 genes were significantly down-regulated after YM155 treatment. Ingenuity pathway analysis (IPA) showed cell death was the highest rated network with 65 focus molecules and the significance score of 44. The IPA analysis also groups the differentially expressed genes into biological mechanisms that are related to cell death, cellular function maintenance, cell morphology, carbohydrate metabolism and cellular growth and proliferation. Death receptor signaling (3.87E-19), TNFR1 signaling, induction of apoptosis by HIV1, apoptosis signaling and molecular mechanisms of cancer came out to be the top four most significant pathways. IPA analysis also showed top molecules up-regulated were BBC3, BIRC3, BIRC8, BNIP1, CASP7, CASP9, CD5, CDKN1A, CEBPG and COL4A3, top molecules down-regulated were ZNF443, UTP11L, TP73, TNFSF10, TNFRSF1B, TNFRSF25, TIAF1, STK17A, SST and SPP1, upstream regulator were NR3C1, TP53, dexamethasone , TNF and Akt.ConclusionsThe present study demonstrates that YM155 treatment resulted in apoptosis and inhibition of cell proliferation of SK-NEP-1cells. YM155 had significant role and little side effect in the treatment of SK-NEP-1 xenograft tumors. Real-time PCR array analysis firstly showed expression profile of genes dyes-regulated after YM155 treatment. IPA analysis also represents new molecule mechanism of YM155 treatment, such as NR3C1 and dexamethasone may be new target of YM155. And our results may provide new clues of molecular mechanism of apoptosis induced by YM155.
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11.4
作者：
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