Transcription and splicing regulation in human umbilical vein endothelial cells under hypoxic stress conditions by exon array.
Transcription and splicing regulation in human umbilical vein endothelial cells under hypoxic stress conditions by exon array.
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外显子阵列在缺氧应激条件下调控人脐静脉内皮细胞的转录和剪接
DOI：
10.1186/1471-2164-10-126
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发表时间：
2009-03-25
期刊：
影响因子：
4.4
通讯作者：
Zhang C
中科院分区：
文献类型：
--
作者：
Hang X;Li P;Li Z;Qu W;Yu Y;Li H;Shen Z;Zheng H;Gao Y;Wu Y;Deng M;Sun Z;Zhang C
应激条件下内皮细胞的存活和凋亡之间的平衡是维持血管完整性和血管稳态的重要细胞过程，也是许多血管疾病发生过程中血管生成的关键。然而，潜在的分子机制在很大程度上仍然未知。尽管转录和选择性剪接在调节应激状态下内皮细胞的基因表达方面都很重要，但这种状态下的调节机制及其相互作用尚未在全基因组基础上进行研究。结果用氯化钴（CoCl2）处理人脐静脉内皮细胞（HUVECs），模拟缺氧，诱导细胞凋亡和选择性剪接反应。细胞凋亡率分析表明，暴露于300 μM cocl2 24小时后，huvec在初始阶段能够平衡细胞凋亡与细胞存活之间的关系。因此，我们使用Affymetrix外显子阵列系统来确定全基因组转录本和外显子水平的差异表达。除了1583个差异表达转录本外，还检测到342个选择性剪接外显子，并根据不同的剪接类型进行分类。16个选择性剪接外显子通过RT-PCR验证。此外，基因本体（Gene Ontology， GO）和蛋白质功能，以及差异表达转录物的蛋白质结构域和途径富集分析，为HUVEC存活和凋亡之间的持续平衡提供了直接证据。重要的是，一个新的分子模块，其中热休克蛋白（HSP）家族发挥重要作用，被发现在模拟缺氧条件下被激活。此外，46%含有应力调节外显子的转录本存在差异表达，表明转录和剪接可能存在组合调控。结论外显子阵列系统能有效地在全基因组范围内分析基因表达和剪接。基于这种方法，我们的数据表明，转录和剪接不仅调节基因表达，还对模拟缺氧条件下HUVECs的生存与凋亡平衡进行组合调控。由于细胞凋亡挑战后的细胞存活在许多血管疾病的发展过程中是血管生成的关键，我们的研究结果可能会促进对内皮细胞在生理和病理条件下多水平基因调控的认识。
BackgroundThe balance between endothelial cell survival and apoptosis during stress is an important cellular process for vessel integrity and vascular homeostasis, and it is also pivotal in angiogenesis during the development of many vascular diseases. However, the underlying molecular mechanisms remain largely unknown. Although both transcription and alternative splicing are important in regulating gene expression in endothelial cells under stress, the regulatory mechanisms underlying this state and their interactions have not yet been studied on a genome-wide basis.ResultsHuman umbilical vein endothelial cells (HUVECs) were treated with cobalt chloride (CoCl2) both to mimic hypoxia and to induce cell apoptosis and alternative splicing responses. Cell apoptosis rate analysis indicated that HUVECs exposed to 300 μM CoCl2for 24 hrs were initially counterbalancing apoptosis with cell survival. We therefore used the Affymetrix exon array system to determine genome-wide transcript- and exon-level differential expression. Other than 1583 differentially expressed transcripts, 342 alternatively spliced exons were detected and classified by different splicing types. Sixteen alternatively spliced exons were validated by RT-PCR. Furthermore, direct evidence for the ongoing balance between HUVEC survival and apoptosis was provided by Gene Ontology (GO) and protein function, as well as protein domain and pathway enrichment analyses of the differentially expressed transcripts. Importantly, a novel molecular module, in which the heat shock protein (HSP) families play a significant role, was found to be activated under mimicked hypoxia conditions. In addition, 46% of the transcripts containing stress-modulated exons were differentially expressed, indicating the possibility of combinatorial regulation of transcription and splicing.ConclusionThe exon array system effectively profiles gene expression and splicing on the genome-wide scale. Based on this approach, our data suggest that transcription and splicing not only regulate gene expression, but also carry out combinational regulation of the balance between survival and apoptosis of HUVECs under mimicked hypoxia conditions. Since cell survival following the apoptotic challenge is pivotal in angiogenesis during the development of many vascular diseases, our results may advance the knowledge of multilevel gene regulation in endothelial cells under physiological and pathological conditions.
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影响因子：
4.5
作者：
Ip, Joanna Y.;Tong, Alan;Lynch, Kristen W.
通讯作者：
Lynch, Kristen W.
影响因子：
7.3
作者：
Cance, William G.;Golubovskaya, Vita M.
通讯作者：
Golubovskaya, Vita M.
DOI：
10.1006/bbrc.2001.5802
发表时间：
2001-11-02
影响因子：
3.1
作者：
Hatayama, T;Yamagishi, N;Sakai, K
通讯作者：
Sakai, K
影响因子：
20.1
作者：
Ji, YS;Xu, Q;Schmedtje, JF
通讯作者：
Schmedtje, JF
影响因子：
20.3
作者：
Birdsey, Graeme M.;Dryden, Nicola H.;Randi, Anna M.
通讯作者：
Randi, Anna M.