Identification of apoptosis-inducing factor in human coronary artery endothelial cells.
Apoptosis has been observed in vascular tissues in response to pro-inflammatory and pro-atherosclerotic stimuli, such as oxidized low density lipoproteins (ox-LDL), angiotensin II (Ang II), and tumor necrosis factor-alpha (TNF-alpha). Apoptosis is believed to be mediated via caspase-dependent pathway. Recently, a 57 kDa molecule, apoptosis-inducing factor (AIF), has been described as a basis for cell injury via a caspase-independent pathway. This study was designed to identify the presence of AIF and the regulation of its gene expression in human coronary artery endothelial cells (HCAECs). Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to determine AIF mRNA and protein expression. Cultured HCAECs were treated with ox-LDL (10-40 microg/ml), angiotensin II (10(-9)-10(-6)M), or TNF-alpha (0.1-10n g/ml). AIF was barely detectable in unstimulated HCAECs; however, treatment with ox-LDL, but not with Ang II or TNF-alpha, significantly increased the expression of AIF in a concentration- and time-dependent manner. DNA sequencing analysis substantiated the existence of AIF in the HCAECs. Treatment of cells with the caspase inhibitor with Z-VAD-fmk did not change ox-LDL-mediated AIF protein expression. Ox-LDL-mediated upregulation of AIF expression was inhibited by actinomycin D, suggesting transcriptional regulation. Further, upon treatment of cells with ox-LDL AIF translocated from mitochondria to the nucleus, as determined by immunocytochemistry. These data suggest that AIF is expressed in HCAECs and is upregulated by ox-LDL.[1]References
- Identification of apoptosis-inducing factor in human coronary artery endothelial cells. Zhang, W., Shokeen, M., Li, D., Mehta, J.L. Biochem. Biophys. Res. Commun. (2003) [Pubmed]
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