Oridonin-induced apoptosis of Jurkat cells and its mechanism

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• 作者：Jiajun Liu ; Renwei Huang ; Dongjun Lin ; Xiangyuan Wu ; Xinyao wu ; Huiling Lu ; Xianglin Pan ; Jun Peng ; Mingquan Li and Qu Lin
• 关键词：Apoptosis ; Caspase ; 3 ; Jurkat cell ; Oridonin ; PARP
• 刊名：Comparative Clinical Pathology
• 出版年：2004
• 出版时间：November 2004
• 年：2004
• 卷：13
• 期：2
• 页码：65-69
• 全文大小：271 KB
• 刊物类别：Medicine
• 刊物主题：Medicine & Public Health
  Pathology
  Hematology
  Oncology
  
• 出版者：Springer London
• ISSN：1618-565X

文摘

Jurkat cells in culture medium in vitro were exposed to different concentrations of oridonin. The proliferation rate of the cells was measured by MTT assay, cell apoptotic rate was detected by flow cytometry, morphology of cell apoptosis was observed by Hoechst 33258 fluorescence staining, DNA fragmentation was assayed by agarose gel electrophoresis, caspase-3 and poly(ADP-ribose) polymerase (PARP) expressions were detected by Western blotting using polyclonal anti-caspase-3 antibody and mouse anti-human PARP monoclonal antibodies, and caspase-3 activity was assayed with a colorimetric assay kit before and after apoptosis had occurred. Oridonin (over 32 mol/l) inhibited the growth of Jurkat cells and caused significant apoptosis. The suppression was both time-dependent and dose-dependent. Marked morphological changes of cell apoptosis, including condensation of chromatin and nuclear fragmentation, were clearly observed by Hoechst 33258 fluorescence staining, as well as by agarose gel electrophoresis. Western blotting showed cleavage of the caspase-3 zymogen protein (32 kDa), with the appearance of its 17 kDa subunit, and a cleaved 89-kDa fragment of 116 kDa PARP was also found, together with a concurrent increase in caspase-3 apoptotic activity. Oridonin can induce apoptosis in Jurkat cells via activation of caspase-3; the results indicate that oridonin might be an important potential anti-leukaemia reagent.