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KMID : 0613820080180040522
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Journal of Life Science
2008 Volume.18 No. 4 p.522 ~ p.529
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Characterization of HtrA2-deficient Mouse Embryonic Fibroblast Cells Based on Morphology and Analysis of their Sensitivity in Response to Cell Death Stimuli
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Lee Sang-Kyu
Nam Min-Kyoung
Kim Goo-Young
Rhim Hyang-Shuk
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Abstract
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High-temperature requirement A2(HtrA2) has been known as a human homologue of bacterial HtrA that has a molecular chaperone function. HtrA2 is mitochondrial serine protease that plays a significant role in regulating the apoptosis; however, the physiological function of HtrA2 still remains elusive. To establish experimental system for the investigation of new insights into the function of HtrA2 in mammalian cells, we first obtained HtrA2+/+ and HtrA2-/- MEF cells lines and identified those cells based on the expression pattern and subcellular localization of HtrA2, using immunoblot and biochemical assays. Additionally, we observed that the morphological characteristics of HtrA2-/- MEF cells are different form those of HtrA2+/+ MEF cells, showing a rounded shape instead of a typical fibroblast-like shape. Growth rate of HtrA2-/- MEF cells was also 1.4-fold higher than that of HtrA2+/+ MEF cells at 36 hours. Furthermore, we verified both MEF cell lines induced caspsase-dependent cell death in response to apoptotic stimuli such as heat shock, staurosporine, and rotenone. The relationship between HtrA2 and heat shock-induced cell death is the first demonstration of the research field of HtrA2. Our study suggests that those MEF cell lines are suitable reagents to further investigate the molecular mechanism by which HtrA2 regulates the balance between cell death and survival.
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KEYWORD
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High-temperature requirement A2, mouse embryonic fibroblast (MEF) cells, cell death, heat shock
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