Microarray analysis of cell cycle gene expression in adult human corneal endothelial cells.
Microarray analysis of cell cycle gene expression in adult human corneal endothelial cells.
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DOI：
10.1371/journal.pone.0094349
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Thuret G
中科院分区：
文献类型：
--
作者：
Ha Thi BM;Campolmi N;He Z;Pipparelli A;Manissolle C;Thuret JY;Piselli S;Forest F;Peoc'h M;Garraud O;Gain P;Thuret G
角膜内皮细胞（EC）形成控制角膜的水合作用并因此控制其透明度的单层。它们在人类中几乎为零的增殖状态在几种常见疾病中是导致不可逆的角膜混浊的细胞池磨损的原因。为了筛选参与细胞周期阻滞的候选基因，我们研究了与体内EC相比被认为诱导不同增殖谱的各种环境下的人EC。将供体角膜（死亡后几小时）、器官培养（OC）角膜、体外融合和非融合原代培养物以及永生化EC系与角膜移植最初几分钟内取出的健康EC进行比较。使用112个细胞周期关键基因的cDNA阵列比较转录谱，并使用Gene Ontology分类进行分析; GenMAPP进行细胞周期调控途径的聚类分析和基因图谱展示。使用qRT-PCR对11个选定的基因验证结果。我们发现了几个转录的蛋白质参与细胞周期阻滞，而不是以前在人类EC报告。早期G1期阻滞效应子和多种DNA损伤诱导的细胞周期阻滞相关转录物在体内被发现，并在OC和体外EC中过度表达。虽然高度增殖，永生化的EC也表现出过度表达的转录涉及细胞周期阻滞。这些新的效应物可能解释了人类成年EC的特征性应激诱导的过早衰老。它们是触发和控制EC增殖的潜在靶点，以期增加储存角膜的细胞库或促进生物工程内皮移植物的大规模EC培养。
Corneal endothelial cells (ECs) form a monolayer that controls the hydration of the cornea and thus its transparency. Their almost nil proliferative status in humans is responsible, in several frequent diseases, for cell pool attrition that leads to irreversible corneal clouding. To screen for candidate genes involved in cell cycle arrest, we studied human ECs subjected to various environments thought to induce different proliferative profiles compared to ECs in vivo. Donor corneas (a few hours after death), organ-cultured (OC) corneas, in vitro confluent and non-confluent primary cultures, and an immortalized EC line were compared to healthy ECs retrieved in the first minutes of corneal grafts. Transcriptional profiles were compared using a cDNA array of 112 key genes of the cell cycle and analysed using Gene Ontology classification; cluster analysis and gene map presentation of the cell cycle regulation pathway were performed by GenMAPP. Results were validated using qRT-PCR on 11 selected genes. We found several transcripts of proteins implicated in cell cycle arrest and not previously reported in human ECs. Early G1-phase arrest effectors and multiple DNA damage-induced cell cycle arrest-associated transcripts were found in vivo and over-represented in OC and in vitro ECs. Though highly proliferative, immortalized ECs also exhibited overexpression of transcripts implicated in cell cycle arrest. These new effectors likely explain the stress-induced premature senescence that characterizes human adult ECs. They are potential targets for triggering and controlling EC proliferation with a view to increasing the cell pool of stored corneas or facilitating mass EC culture for bioengineered endothelial grafts.
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影响因子：
2
作者：
Bittel, Douglas C.;Kibiryeva, Nataliya;Butler, Merlin G.
通讯作者：
Butler, Merlin G.
影响因子：
3.8
作者：
Bryan, BA;Dyson, OF;Akula, SM
通讯作者：
Akula, SM
影响因子：
2
作者：
Bayyoud, Tarek;Thaler, Sebastian;Yoeruek, Efdal
通讯作者：
Yoeruek, Efdal
影响因子：
6.2
作者：
Gentile, Agustina;Ditt, Renata F.;Menossi, Marcelo
通讯作者：
Menossi, Marcelo
影响因子：
3.4
作者：
IKEBE, H;TAKAMATSU, T;FUJITA, S
通讯作者：
FUJITA, S