Study on the expression of IkappaB-alpha protein in TNF-alpha induced apoptosis of U937 cells.
OBJECTIVE: To investigate the TNF-alpha induced apoptosis of U937 cells, the expression, degradation and subcellular localization of IkappaB-alpha, and its degradation mechanism. METHOD: Changes and subcellular loca-lization of IkappaB-alpha were observed by fluorescence microscopy, expression and degradation of IkappaB-alpha protein with N-tosyl-L-phenylalanylchloromethyl ketone (TPCK protease inhibitor) blocking test and apoptosis of U937 cell by flow cytometry. RESULTS: (1) immunolfluorescence assay showed that IkappaB-alpha localized in cytoplasm only. (2) The level of IkappaB-alpha protein was downregulated after TNF-alpha stimulation, flow cytometry also confirmed the downregulation. (3) The downregulation of IkappaB-alpha protein levels in TNF-alpha induced apoptosis was partially inhibited by TPCK. (4) The apoptosis rate of U937 cells induced by TNF-alpha was (60.73 +/- 1.61)%. CONCLUSION: (1) Degradation of IkappaB-alpha protein during TNF-alpha induced apoptosis of U937 cells suggested the activation of NF-kappaB. (2) TPCK sensitive protease plays an important role in the degradation of IkappaB-alpha protein. (3) TPCK sensitive protease also involved in the apoptosis of U937 cells induced by TNF-alpha.[1]References
- Study on the expression of IkappaB-alpha protein in TNF-alpha induced apoptosis of U937 cells. Chen, W., Peng, G., Tang, A., Wang, K., Zhou, M., Wang, L. Zhonghua Xue Ye Xue Za Zhi (2002) [Pubmed]
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