Abstract
Expressions of the effector molecules of Fas-mediated apoptosis in primary cultured salivary gland epithelial cells (SGEC) of primary Sjögren’s syndrome (pSS) remain to be clarified. We focused on Fas-mediated caspase cleavage compared to tumor necrosis factor-related apoptosis inducing ligand (TRAIL)-mediated apoptosis. Induction of apoptosis was performed by anti-Fas antibody coupled with PI3K inhibitor, or TRAIL. Activation of caspases, cytochrome C, and apoptotic protease activating factor-1 (Apaf-1) was determined by western blotting or immunofluorescence observed by confocal microscopy. Fas-mediated apoptosis and activation of caspase 3/8 were induced in the presence of LY294002. TRAIL-induced apoptosis in SGEC, which was stronger than that induced by anti-Fas antibody. TRAIL-induced caspase 9 cleavage accompanied by activation of cytochrome C and Apaf-1 were not mediated by anti-Fas antibody. Our results suggest that death receptor-dependent apoptosis in primary cultured SGEC is regulated by the engagement of type II cells in pSS.
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Abbreviations
- Apaf-1:
-
Apoptotic protease activating factor-1
- EGF:
-
Epidermal growth factor
- FITC:
-
Fluorescein isothiocyanate
- LSG:
-
Labial salivary glands
- PBS:
-
Phosphate-buffered saline
- PI3K:
-
Phosphatidylinositol 3-kinase
- pSS:
-
Primary Sjögren’s syndrome
- SGEC:
-
Salivary gland epithelial cells
- TRAIL:
-
Tumor necrosis factor-related apoptosis inducing ligand
- TRITC:
-
Tetramethyl rhodamine isothiocyanate
- TUNEL:
-
Terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling
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Acknowledgments
The authors would like to thank Dr. Noriyoshi Ogawa from the Third Department of Internal Medicine, Hamamatsu Medical University, for advice regarding the primary cell culture of epithelial cells obtained from minor salivary gland biopsy.
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Nakamura, H., Kawakami, A., Iwamoto, N. et al. Rapid and significant induction of TRAIL-mediated type II cells in apoptosis of primary salivary epithelial cells in primary Sjögren’s syndrome. Apoptosis 13, 1322–1330 (2008). https://doi.org/10.1007/s10495-008-0261-2
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DOI: https://doi.org/10.1007/s10495-008-0261-2