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CHANGES IN GENE EXPRESSION DURING DIFFERENTIATION OF CULTURED HUMAN PRIMARY BRONCHIAL EPITHELIAL CELLS
Citation:
ROSS, A., L. A. DAILEY, AND R. B. DEVLIN. CHANGES IN GENE EXPRESSION DURING DIFFERENTIATION OF CULTURED HUMAN PRIMARY BRONCHIAL EPITHELIAL CELLS. Presented at American Thoracic Society International Conference, San Diego, CA, May 19 - 24, 2006.
Description:
Primary airway epithelial cell cultures are a useful tool for the in vitro study of normal bronchial cell differentiation and function, airway disease mechanisms, and pathogens and toxin response. Growth of these cells at an air-liquid interface for several days results in the formation of a pseudostratified epithelium composed of ciliated and mucus-secreting cells. While these cultured cells display many functional and morphological characteristics of the airway epithelium, less is known about them at the level of gene expression. We performed microarray analysis to understand the changes that occur to these cells in culture, and to identify genes and pathways involved in airway epithelial cell differentiation. Cells from three donors were cultured and harvested at 10 timepoints over a 28 day period. RNA from the cells was profiled using Affymetrix HG-U133 Plus arrays. We identified over 3000 genes that displayed a statistically significant 2-fold or greater change in expression during the time course. Of the genes showing the largest increases, many are known to be expressed in the respiratory tract (PLUNC, SCGB3A1, C9orf24, FOXJ1), encode proteins associated with cilia or axonemes (SPAG6, AKAP14, ROPN1L , TUBA3) or mucus production (MUC5B); however, many novel genes were also identified. Gene networks were generated using Ingenuity Pathways Analysis (Ingenuity Systems), which identified potential roles for TGFB1, TP53, and MYC during this process. Additional gene ontology and pathway analyses have identified unexpected classes of genes that are changed over the course of the culture period. Thus, this work has provided a wealth of information about gene expression during differentiation of airway epithelial cells in vitro, and provides a useful resource for researchers who use this culture system. This abstract does not necessarily represent EPA policy.