Microarray Analysis of Cell Cycle Gene Expression in Adult Human Corneal Endothelial Cells
| Autor: | Gilles Thuret, Zhiguo He, Binh Minh Ha Thi, Jean-Yves Thuret, Olivier Garraud, Fabien Forest, Simone Piselli, Michel Peoc'h, Nelly Campolmi, C. Manissolle, A. Pipparelli, Philippe Gain |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Cell division
Microarrays Cell lcsh:Medicine Genetic Networks Medicine and Health Sciences lcsh:Science Cellular Senescence Oligonucleotide Array Sequence Analysis Aged 80 and over Multidisciplinary biology Gene Ontologies Cell Cycle Endothelium Corneal Genomics Cell cycle Middle Aged Cell Cycle Gene Sensory Systems Cyclin-Dependent Kinases Cell biology medicine.anatomical_structure Bioassays and Physiological Analysis Corneal Disorders Cell aging Transcriptome Analysis Cell Division Research Article Signal Transduction Adult Surgical and Invasive Medical Procedures Research and Analysis Methods Cell Line Organ Culture Techniques Cyclin-dependent kinase Cyclins medicine Genetics Animals Humans Molecular Biology Techniques Molecular Biology Aged Transplantation Microarray analysis techniques Gene Expression Profiling lcsh:R Gene Mapping Biology and Life Sciences Computational Biology Genome Analysis Molecular biology Ophthalmology Cell culture biology.protein lcsh:Q Genome Expression Analysis Neuroscience |
| Zdroj: | PLoS ONE PLoS ONE, Vol 9, Iss 4, p e94349 (2014) |
| ISSN: | 1932-6203 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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