Gene induction during differentiation of human pulmonary type II cells in vitro.
Mature alveolar type II cells that produce pulmonary surfactant are essential for adaptation to extrauterine life. We profiled gene expression in human fetal lung epithelial cells cultured in serum-free medium containing dexamethasone and cyclic AMP, a treatment that induces differentiation of type II cells. Microarray analysis identified 388 genes that were induced > 1.5-fold by 72 h of hormone treatment. Induced genes represented all categories of molecular function and subcellular location, with increased frequency in the categories of ionic channel, cell adhesion, surface film, lysosome, extracellular matrix, and basement membrane. In time-course experiments, self-organizing map analysis identified a cluster of 17 genes that were slowly but highly induced (5- to approximately 190-fold) and represented four functional categories: surfactant-related (SFTPC, SFTPA, PGC, SFTPB, LAMP3, LPL), regulatory (WIF2, IGF2, IL1RL1, NR4A2, HIF3A), metabolic (MAOA, ADH1B, SEPP1), and transport (SCNN1A, CLDN18, AQP4). Induction of both mRNA and protein for these genes, which included nine newly identified regulated genes, was confirmed, and cellular localization was determined in both fetal and postnatal tissue. Induction of lysosomal-associated membrane protein 3 required both hormones, and expression was localized to limiting membranes of lamellar bodies. Hormone-induced differentiation of human type II cells is associated with genome-wide increased expression of genes with diverse functions.
Wade
Kelly C
KC
Department of Pediatrics, Division of Neonatology, Children's Hospital of Philadelphia, Philadelphia, PA, USA.
Guttentag
Susan H
SH
Gonzales
Linda W
LW
Maschhoff
Kathryn L
KL
Gonzales
John
J
Kolla
Venkatadri
V
Singhal
Sunil
S
Ballard
Philip L
PL
D015124:8-Bromo Cyclic Adenosine Monophosphate; D002454:Cell Differentiation; D002478:Cells, Cultured; D003907:Dexamethasone; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D064429:Fatty Acid Synthases / Q000235:genetics / Q000378:metabolism; D005326:Fetal Proteins / Q000235:genetics / Q000378:metabolism; D005333:Fetus; D020869:Gene Expression Profiling / Q000379:methods; D018507:Gene Expression Regulation, Developmental; D005991:Glycerol Kinase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008071:Lipoprotein Lipase / Q000235:genetics / Q000378:metabolism; D008168:Lung / Q000378:metabolism*; D051907:Lysosomal Membrane Proteins / Q000235:genetics; D010743:Phospholipids / Q000096:biosynthesis; D012333:RNA, Messenger / Q000032:analysis; D013347:Subcellular Fractions / Q000378:metabolism; D013997:Time Factors; D015533:Transcriptional Activation; D017855:Transferases (Other Substituted Phosphate Groups) / Q000235:genetics / Q000378:metabolism
D005326:Fetal Proteins; D051907:Lysosomal Membrane Proteins; D010743:Phospholipids; D012333:RNA, Messenger; D015124:8-Bromo Cyclic Adenosine Monophosphate; D003907:Dexamethasone; D064429:Fatty Acid Synthases; D005991:Glycerol Kinase; D017855:Transferases (Other Substituted Phosphate Groups); C020676:CDP-diacylglycerol-glycerol-3-phosphate 3-phosphatidyltransferase; D008071:Lipoprotein Lipase