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首页> 外文期刊>Comparative and functional genomics >Early gene expression in wounded human keratinocytes revealed by DNA microarray analysis
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Early gene expression in wounded human keratinocytes revealed by DNA microarray analysis

机译：DNA芯片分析揭示了受伤的人类角质形成细胞中的早期基因表达

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Wound healing involves several steps: spreading of the cells, migration and proliferation. We have profiled gene expression during the early events of wound healing in normal human kerationcytes with a home-made DNA microarray containing about 1000 relevant human probes. An original wounding machine was used, that allows the wounding of up to 40% of the surface of a confluent monolayer of cultured cells grown on a Petri dish (compared with 5% with a classical "scratch' method). The two aims of the present study were: (a) to validate a limited number of genes by comparing the expression levels obtained with this technique with those found in the literature; (b) to combine the use of the wounding machine with DNA microarray analysis for large-scale detection of the molecular events triggered during the early stages of the wound-healing process. The time-courses of RNA expression observed at 0.5, 1.5, 3, 6 and 15 h after wounding for genes such as c-Fos, c-Jun, Egr1, the plasminogen activator PLAU (uPA) and the signal transducer and transcription activator STAT3, were consistent with previously published data. This suggests that our methodologies are able to perform quantitative measurement of gene expression. Transcripts encoding two zinc finger proteins, ZFP26 and ZNF161, and the tumour necrosis factor α-induced protein TNFAIP3, were also overexpressed after wounding. The role of the p38 mitogen-activated protein kinase (p38MAPK) in wounding. The role of the p38 mitogen-activated protein kinase (p38MAPK) in wound healing was shown after the inhibition of p38 by SB203580, but our results also suggest the existence of surrogate activating pathways.
机译：伤口愈合涉及几个步骤：细胞扩散，迁移和增殖。我们已用包含约1000种相关人类探针的自制DNA微阵列分析了正常人类克星细胞在伤口愈合的早期事件中的基因表达。使用原始的伤口机，最多可以伤口在培养皿上生长的培养细胞汇合单层表面的40％（相对于传统的“刮擦”方法为5％）。当前的研究是：（a）通过比较用此技术获得的表达水平与文献中发现的表达水平来验证有限的基因;（b）将伤口机的使用与DNA微阵列分析相结合以进行大规模检测在伤口愈合过程的早期触发的分子事件的发生，在受伤后0.5、1.5、3、6和15 h观察到的c-Fos，c-Jun，Egr1等基因RNA表达的时程纤溶酶原激活物PLAU（uPA）以及信号转导子和转录激活子STAT3与以前发表的数据一致，这表明我们的方法能够对基因表达进行定量测量。编码两个锌指蛋白的转录本ZFP26和ZNF161以及肿瘤坏死因子α诱导的蛋白TNFAIP3在受伤后也过表达。 p38丝裂原活化蛋白激酶（p38MAPK）在伤口中的作用。 SB203580抑制p38后，显示了p38促分裂原活化蛋白激酶（p38MAPK）在伤口愈合中的作用，但我们的结果也表明存在替代激活途径。

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