J Korean Ophthalmol Soc.  2003 Nov;44(11):2615-2626.

Profiling of Differentially Expressed Genes in Human Polymorphonuclear Leukocyte on Human Amniotic Membrane

Affiliations
  • 1Department of Ophthalmology, Kangdong Sacred Heart Hospital, Hallym University College of Medicine, Korea. thc@hallym.or.kr
  • 2Dr. Lee's Eye Clinic, Korea.

Abstract

PURPOSE
To identify genes that showed altered expression between human polymorphonuclear (PMN) cell cultured on plastic and on amniotic membrane by the technique of differential hybridization of two Altas(TM) Human cDNA expression array. METHODS: 32P-labeled complimentary DNA probes derived from RNA of either human polymorphonuclear leukocyte cultured on plastic and cultured on amniotic membrane were hybridized to two identical human cDNA expression array membranes containing 588 known genes. RESULTS: Of the total 588 genes, 130 genes were up- or down-regulated. 50 up-regulated and 80 down-regulated genes were identified in polymorphonuclear leukocyte cultured on amniotic membrane compared with control. After different signal intensity was normalized more than 4000 by Atlas Image(TM) 1.0 Software, 19 genes were up-regulated and 36 genes down-regulated. CONCLUSIONS: Genes associated with the process of apoptosis, DNA synthesis and repair were down-regulated in PMN cultured on AM and genes associated with DNA binding protein, transcription factor were altered. Cell-cell communication factors including TGF-beta, PDGF-A, RANTES, MRP-14, oncostatin M, MIP-2 alpha were significantly down-regulated and cell surface antigen CD11a (LFA-1) was down-regulated, suggesting that AM can suppress the inflammatory reaction mediated by adhesion molecule, inflammatory, proinflammatory cytokines and chemokines.

Keyword

Amniotic membrane; cDNA microarray; Inflammation; Polymorphonuclear cell

MeSH Terms

Amnion*
Antigens, Surface
Apoptosis
Chemokine CCL5
Chemokines
Cytokines
DNA
DNA Probes
DNA, Complementary
DNA-Binding Proteins
Humans*
Inflammation
Membranes
Neutrophils*
Oligonucleotide Array Sequence Analysis
Oncostatin M
Plastics
RNA
Transcription Factors
Transforming Growth Factor beta
Antigens, Surface
Chemokine CCL5
Chemokines
Cytokines
DNA
DNA Probes
DNA, Complementary
DNA-Binding Proteins
Oncostatin M
Plastics
RNA
Transcription Factors
Transforming Growth Factor beta
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