Enhanced Expression of Membrane-Associated Sialidase Neu3 Decreases GD3 and Increases GM3 on the Surface of Jurkat Cells during Etoposide-Induced Apoptosis(Molecular and Cell Biology)

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概要

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• We previously reported that, in Jurkat human T cells, the topoisomerase II inhibitor etoposide enhances sialidase activity and reduces cell surface sialic acid levels at an early stage of apoptosis and that the decreases in sialic acid are suppressed by the sialidase inhibitor 2,3-dehydro-2-deoxy-N-acetylneuraminic acid [Azuma Y., et al., Glycoconj. J., 17, 301-306 (2000)]. In the current studies, we treated Jurkat cells with etoposide and exam- ined the changes in the cell surface levels of gangliosides GM1, GM2, GM3, GD1a, and GD3 at physiological pH using anti-ganglioside antibodies. We also examined the sialidase activity on the cell surface using 4-methylum-belliferyl N-acetylneuraminic acid and measured the mRNA expression of the plasma membrane-associated sialidase Neu3 and the lysozomal Neul using real-time PCR. We found an increase in GM3 and a decrease in GD3 during the early stage (4 h) of etoposide-induced apoptosis that preceded the increase in cell surface exposure ofphosphatidylserine (4 to 6 h). The caspase 3 inhibitor acetyl-Asp-Glu-Val-Asp-aldehyde significantly suppressedchanges in GM3 and GD3 and blocked the enhanced cell surface sialidase activity. Furthermore, etoposidecaused a gradual up-regulation of Neu3 mRNA expression but not Neul mRNA expression. Enhanced Neu3mRNA expression was suppressed in the presence of caspase 3 inhibitor. These results indicate that Neu3 is up-regulated in Jurkat cells undergoing etoposide-induced apoptosis through intracellular signaling events down-stream of caspase 3 activation and that enhanced Neu3 activity is closely related to the changes of cell surface ganglioside composition.

• 2007-09-01

著者

• SATO Hirotaka

  Department of Electrical and Communication Engineering, Graduate School of Engineering, Tohoku Unive

• Sato Hirotaka

  Department Of Clinical Chemistry School Of Pharmaceutical Sciences Toho University

• HIGAI Koji

  Department of Clinical Chemistry, Faculty of Pharmaceutical Sciences, Toho University

• MATSUMOTO Kojiro

  Department of Clinical Chemistry, Faculty of Pharmaceutical Sciences, Toho University

• Higai Koji

  Department Of Clinical Chemistry School Of Pharmaceutical Sciences Toho University

• Azuma Yutaro

  Department of Clinical Chemistry, Faculty of Pharmaceutical Sciences, Toho University

• Azuma Y

  Dep. Of Clinical Chemistry Fac. Of Pharmaceutical Sciences Toho Univ.

• Azuma Yutaro

  Department Of Biochemistry Tokyo Dental College

• Matsumoto Kojiro

  Dep. Of Clinical Chemistry Fac. Of Pharmaceutical Sciences Toho Univ.

• Matsumoto Kojiro

  Department Of Clinical Chemistry Faculty Of Pharmaceutical Sciences Toho University

• Higai Koji

  Department of Clinical Chemistry, School of Pharmaceutical Sciences, Toho University

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