PMID- 19279691 OWN - NLM STAT- MEDLINE DCOM- 20090512 LR - 20211020 IS - 1090-0535 (Electronic) IS - 1090-0535 (Linking) VI - 15 DP - 2009 TI - Alterations in gene expression induced by cyclic mechanical stress in trabecular meshwork cells. PG - 534-44 AB - PURPOSE: To investigate the changes in gene expression induced by cyclic mechanical stress (CMS) in trabecular meshwork (TM) cells. METHODS: Human TM cultures from three donors were plated on type I collagen-coated flexible silicone bottom plates and subjected to 15% stretching, one cycle per second for 6 h. Non-stressed parallel control cultures were incubated under the same conditions in the absence of CMS. Total RNA from each culture was amplified (1 round of amplification) and hybridized to Operon Human Oligo Arrays version 3.0 (35 K probes). Differences in gene expression induced by CMS were analyzed using Genespring 7.2. quantitative polymerase chain reaction (Q-PCR) was used to confirm changes in the expressions of 12 selected genes. The effects of chemical inhibitors for p38, ERK (extracellular signal-regulated kinase), JNK (Jun N-terminal kinase), PKA (protein kinase A), PI3K (phosphoinositide 3-kinase), and P2 (purinergic 2) receptors on the induction of MMP3 (matrix metalloproteinase 3), HSP70 (heat shock protein 70), ECSM1 (endothelial cell specific molecule 1), BMP2 (bone morphogenetic protein 2), VEGFC (vascular endothelial growth factor C), and IL-8 (interleukin 8) were evaluated in porcine TM cells subjected to the same regime of CMS as that used in human cells. RESULTS: CMS induced extensive gene expression changes (664 genes, p < or = 0.05) twofold or higher in cultured TM cells. Many of these changes were related to extracellular matrix (ECM) synthesis and remodeling including the upregulation of two metalloproteinases (MMP3 and MMP10). Cytoskeleton and cell adhesion genes were also affected by CMS as well as genes known to be involved in cellular protection against stress including several members of the HSP70 family. Inhibition of PI3K/AKT and P2 receptors pathways significantly reduced the induction of MMP3 and IL-8 whereas the inhibition of the PKA/cAMP pathway decreased ECSM1 and BMP2. CONCLUSIONS: CMS activated many genes that could influence the aqueous humor outflow facility, specifically genes involved in ECM synthesis and remodeling (e.g. MMPs), cytoskeletal organization, and cell adhesion. Induction of MMP3 has the potential to increase the aqueous humor outflow facility and could be part of a homeostatic mechanism involved in the maintenance of normal intraocular pressure (IOP) levels. Other observed changes are more likely to be related to general cellular responses to stress (e.g., HSP70, ECSM1, and BMP2). Although these latter changes may initially help to repair mechanical damage, some of them such as the induction of BMP2 could eventually increase tissue rigidity and compromise the ability of the TM to maintain normal levels of outflow resistance. FAU - Luna, Coralia AU - Luna C AD - Department of Ophthalmology, Duke University, Durham, NC 27710, USA. FAU - Li, Guorong AU - Li G FAU - Liton, Paloma B AU - Liton PB FAU - Epstein, David L AU - Epstein DL FAU - Gonzalez, Pedro AU - Gonzalez P LA - eng GR - P30 EY005722/EY/NEI NIH HHS/United States GR - EY05722/EY/NEI NIH HHS/United States GR - EY01894/EY/NEI NIH HHS/United States GR - R01 EY016228/EY/NEI NIH HHS/United States GR - R01 EY001894/EY/NEI NIH HHS/United States GR - EY016228/EY/NEI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20090311 PL - United States TA - Mol Vis JT - Molecular vision JID - 9605351 RN - 0 (Cell Adhesion Molecules) RN - 0 (Cytoskeletal Proteins) RN - 0 (Enzyme Inhibitors) RN - 0 (Extracellular Matrix Proteins) RN - 0 (HSP70 Heat-Shock Proteins) RN - 0 (HSPA7 protein, human) RN - EC 3.4.24.17 (MMP3 protein, human) RN - EC 3.4.24.17 (Matrix Metalloproteinase 3) RN - EC 3.4.24.22 (MMP10 protein, human) RN - EC 3.4.24.22 (Matrix Metalloproteinase 10) SB - IM MH - Adolescent MH - Adult MH - Animals MH - Aqueous Humor/metabolism MH - Cell Adhesion Molecules/biosynthesis/genetics MH - Cell Culture Techniques MH - Cell Survival MH - Cytoprotection/genetics MH - Cytoskeletal Proteins/biosynthesis/genetics MH - Enzyme Inhibitors/pharmacology MH - Extracellular Matrix Proteins/biosynthesis/genetics MH - *Gene Expression MH - Gene Expression Profiling MH - HSP70 Heat-Shock Proteins/biosynthesis/genetics MH - Humans MH - MAP Kinase Signaling System/drug effects MH - Matrix Metalloproteinase 10/biosynthesis/genetics MH - Matrix Metalloproteinase 3/biosynthesis/genetics MH - *Stress, Mechanical MH - Swine MH - *Trabecular Meshwork/cytology/physiology PMC - PMC2654047 EDAT- 2009/03/13 09:00 MHDA- 2009/05/13 09:00 PMCR- 2009/01/01 CRDT- 2009/03/13 09:00 PHST- 2008/12/08 00:00 [received] PHST- 2009/02/16 00:00 [accepted] PHST- 2009/03/13 09:00 [entrez] PHST- 2009/03/13 09:00 [pubmed] PHST- 2009/05/13 09:00 [medline] PHST- 2009/01/01 00:00 [pmc-release] AID - 54 [pii] AID - 2008MOLVIS0395 [pii] PST - ppublish SO - Mol Vis. 2009;15:534-44. Epub 2009 Mar 11.