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Optimal Lentivirus Production and Cell Culture Conditions Necessary to Successfully Transduce Primary Human Bronchial Epithelial Cells
Published on: July 22, 2016
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Human bronchial epithelial cells express and secrete MMP-12
Mark C Lavigne1, Paresh Thakker, Jason Gunn
1Wyeth Research, Cardiovascular and Metabolic Diseases, Cambridge, MA 02140, USA.
Biochemical and Biophysical Research Communications
|October 12, 2004
Summary
Normal human bronchial epithelial cells (NHBECs) express and secrete matrix metalloproteinase-12 (MMP-12). Tumor necrosis factor-alpha (TNF-alpha) significantly increases MMP-12 gene expression in NHBECs.
Area of Science:
- Pulmonary Medicine
- Molecular Biology
- Cell Biology
Background:
- Matrix metalloproteinases (MMPs) degrade extracellular matrix proteins.
- MMP activity is implicated in alveolar enlargement in chronic obstructive pulmonary disease (COPD) and pulmonary tissue remodeling in chronic asthma.
Purpose of the Study:
- To investigate the expression and secretion of MMP-12 by normal human bronchial epithelial cells (NHBECs).
- To determine the regulatory effects of tumor necrosis factor-alpha (TNF-alpha), epidermal growth factor (EGF), and interferon gamma (IFN-gamma) on MMP-12 gene expression in NHBECs.
Main Methods:
- Reverse transcription-polymerase chain reaction (RT-PCR) to analyze MMP-12 mRNA levels.
- Western blot analysis to detect MMP-12 protein in cell culture media.
- Stimulation of differentiated NHBEC cultures with TNF-alpha, EGF, and IFN-gamma, individually and in combination.
Main Results:
- MMP-12 mRNA was detected in unstimulated differentiated NHBECs.
- TNF-alpha, alone or in combination with EGF or IFN-gamma, significantly increased MMP-12 mRNA levels within 6 hours.
- EGF and IFN-gamma alone did not alter MMP-12 mRNA abundance.
- MMP-12 protein was confirmed in the conditioned media of differentiated NHBECs.
Conclusions:
- The bronchial epithelium is a potential source of MMP-12, contributing to elastolytic activity in COPD.
- MMP-12 secreted by bronchial epithelial cells may play a role in pulmonary tissue remodeling associated with chronic asthma.
- TNF-alpha is a key regulator of MMP-12 gene expression in the bronchial epithelium.
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