Reverse transcription-polymerase chain reaction (RT-PCR) phenotypic analysis of cell cultures of human tracheal epithelium, tracheobronchial glands, and lung carcinomas.
In order to identify expression of RNA transcripts for a number of important tracheobronchial cell products and molecules, we developed simple reverse transcription-polymerase chain reaction (RT-PCR) assays. Assays included the RNA for two apomucins (MUC1 and MUC2), secretory component, secretory leukocyte inhibitor protein, lysozyme, lactoferrin, 15-lipoxygenase, and the cystic fibrosis transmembrane conductance regulator. We tested RNA of normal and neoplastic origin. Sources of normal tissue included human tracheal surface epithelial cells and tracheobronchial submucosal tissues, acutely isolated human tracheal surface epithelial and tracheobronchial gland acini, and confluent cultures of human tracheal epithelial and tracheobronchial gland cells. Sources of neoplastic tissue included cell lines of non-small cell carcinomas of the lung. RNA expression was correlated with protein expression as assessed by immunocytochemistry. Tracheal surface epithelial tissues, isolated cells and cultures, and tracheobronchial submucosal tissues expressed RNA transcripts for all of the RNA transcripts assayed. Isolated gland acini and cultured gland cells expressed all RNA transcripts except 15-lipoxygenase. Expression of RNA transcripts by non-small cell lung carcinomas was heterogeneous and not necessarily influenced by histopathologic type. In most instances, RNA expression predicted expression of immunocytochemically detectable protein. These RT-PCR assays are useful for characterizing the molecular phenotype of cell cultures derived from normal or neoplastic airway epithelium and for establishing the potential of cultured cells for functional studies.
Finkbeiner
W E
WE
Department of Pathology, University of California, San Francisco 94143-0506.
Carrier
S D
SD
Teresi
C E
CE
D001093:Arachidonate 15-Lipoxygenase / Q000378:metabolism; D001483:Base Sequence; D002277:Carcinoma / Q000378:metabolism*; D002478:Cells, Cultured; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D017931:DNA Primers / Q000737:chemistry; D004848:Epithelium / Q000378:metabolism; D015870:Gene Expression; D007781:Lactoferrin / Q000378:metabolism; D008175:Lung Neoplasms / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D009077:Mucins / Q000378:metabolism; D009113:Muramidase / Q000378:metabolism; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods*; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; D012333:RNA, Messenger / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase; D012635:Secretory Component / Q000378:metabolism; D015842:Serine Proteinase Inhibitors / Q000378:metabolism; D014132:Trachea / Q000378:metabolism*
D017931:DNA Primers; D008565:Membrane Proteins; D009077:Mucins; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; D012333:RNA, Messenger; D012635:Secretory Component; D015842:Serine Proteinase Inhibitors; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D001093:Arachidonate 15-Lipoxygenase; D012194:RNA-Directed DNA Polymerase; D009113:Muramidase; D007781:Lactoferrin