Gene expression profile of B16(F10) murine melanoma cells exposed to hypoxic conditions in vitro
Gene expression profile of B16(F10) murine melanoma cells exposed to hypoxic conditions in vitro
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DOI：
10.3727/000000006783991818
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发表时间：
2006-01-01
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影响因子：
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通讯作者：
Sochanik, Aleksander
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文献类型：
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作者：
Olbryt, Magdalena;Jarzab, Michal;Sochanik, Aleksander
缺氧是肿瘤微环境的一个重要特征，对细胞产生深远的影响，并有助于癌症的进展。以前的研究已经建立了各种细胞系之间的缺氧反应的实质性差异。研究黑色素瘤细胞中的这种现象有助于更好地理解细胞系特异性缺氧反应，并可能指出新的缺氧调节基因。我们研究了B16（F10）小鼠黑色素瘤细胞培养24小时缺氧（标称1%O-2，15个样品，包括控制）和缺氧模拟条件（氯化钴，100或200 μ M，6个样品，包括控制）下的转录活性。使用MG-U 74 Av 2寡核苷酸微阵列分析基因表达谱。数据分析显示2541个探针组（FDR < 5%）用于1%氧实验，364个探针组（FDR < 5%）用于氯化钴实验，其显示表达水平的差异。通过严格的Family-Wise错误率估计对低氧调节基因（真低氧，1%O-2）的分析表明454个显著改变的转录本（p < 0.05）。上调最多的基因是Lgals 3、Selenbp 1、Nppb（增加超过10倍）。我们观察到调节糖酵解（Pfkp、Hk 2、Aldo 3、Eno 2）、凋亡（Bnip 3、Bnip 31、Cdkn 1a）、转录（Bhlhb 2、Sap 30、Atf 3、Mxi 1）、血管生成（Vegfa、Adm、Anxa 2、Ctgl）、粘附（Pkp 2、Itga 4、Mcam）、迁移（Cnn 2、Tmsb 4x）和其他过程的基因的表达水平存在显著差异。真正的缺氧和模拟缺氧诱导HIF-1调节基因。然而，无监督分析（奇异值分解）揭示了这两个实验条件之间的基因表达的明显差异。与缺氧相反，氯化钴引起基因表达的抑制而不是刺激，特别是关于与增殖、免疫反应、DNA修复和黑色素生物合成相关的转录物。
Hypoxia is an important feature of tumor microenvironment, exerting far-reaching effects on cells and contributing to cancer progression. Previous studies have established substantial differences in hypoxia response between various cell lines. Investigating this phenomenon in melanoma cells contributes to a better understanding of cell lineage-specific hypoxia response and could point out novel hypoxia-regulated genes. We investigated transcriptional activity of B16(F10) murine melanoma cells cultured for 24 h under hypoxic (nominal 1% O-2, 15 samples including controls) and hypoxia-mimicking conditions (cobalt chloride, 100 or 200 mu M, 6 samples including controls). Gene expression profiles were analyzed using MG-U74Av2 oligonucleotide microarrays. Data analysis revealed 2541 probesets (FDR < 5%) for 1% oxygen experiment and 364 probesets (FDR < 5%) for cobalt chloride, which showed differences in expression levels. Analysis of hypoxia-regulated genes (true hypoxia, 1% O-2) by stringent Family-Wise Error Rate estimation indicated 454 significantly changed transcripts (p < 0.05). The most upregulated genes were Lgals3, Selenbp1, Nppb (more than ten-fold increase). We observed significant differences in expression levels of genes regulating glycolysis (Pfkp, Hk2, Aldo3, Eno2), apoptosis (Bnip3, Bnip31, Cdkn1a), transcription (Bhlhb2, Sap30, Atf3, Mxi1), angiogenesis (Vegfa, Adm, Anxa2, Ctgl), adhesion (Pkp2, Itga4, Mcam), migration (Cnn2, Tmsb4x), and other processes. Both true hypoxia and hypoxia mimicry induced HIF-1-regulated genes. However, unsupervised analysis (Singular Value Decomposition) revealed distinct differences in gene expression between these two experimental conditions. Contrary to hypoxia, cobalt chloride caused suppression of gene expression rather than stimulation, especially concerning transcripts related to proliferation, immune response, DNA repair, and melanin biosynthesis.