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Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/30690
標題: Induction of apoptosis in SF-21 cell line through caspase pathway by Nomuraea rileyi cultured fluid
綠殭菌菌液藉由caspase 途徑誘發SF-21 細胞產生細胞凋亡
作者: 吳孟學
Meng-ShiueWu
關鍵字: http://etds.lib.nchu.edu.tw/etdservice/view_metadata?etdun=U0005-0908200711115300;綠殭菌;SF-21 細胞株;細胞凋亡;caspase
出版社: 昆蟲學系所
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摘要: 
本研究由細胞型態的改變及DNA-laddering 等現象確認綠殭菌
(Nomuraea rileyi) 培養液能誘發鱗翅目細胞株SF-21 產生細胞凋亡，進一步以TUNEL 染色法確認細胞內DNA 發生斷裂，並發現綠殭菌培養液誘發SF-21 細胞產生細胞凋亡的能力具有劑量依變反應。
Caspase 抑制劑z-VAD-fmk 能阻斷細胞凋亡的發生，說明此細胞凋亡現象乃以caspase 為其調控的中心。在以綠殭菌培養液處理SF-21 細胞後，能偵測到sf-caspase-1 受質之胺基酸序列DEVD 被切割的訊號；經加入抑制劑z-DEVD-fmk 後，發現能阻斷細胞凋亡的發生，證明sf-caspase-1 參與此細胞凋亡現象。而同時加入caspase-8 抑制劑，z-IETD-fmk 及caspase-9 抑制劑，z-LEHD-fmk，與單獨加入其中之一；比較之下，同時加入兩種抑制劑具有更佳的抑制細胞凋亡效果，推測綠殭菌培養液引發之SF-21 細胞細胞凋亡現象，係同時以兩個不同的caspase 活化路徑進行。而在此細胞凋亡過程中偵測到caspase-9 受質之胺基酸序列LEHD 被切割的訊號，因此推測SF-21 細胞之initiator caspase 可能具有與哺乳動物caspase-9 類似之活性。

In this study, we identified that apoptosis in SF-21 cells could be induced by cultured fluid of the entomopathogenic fungus, Nomuraea rileyi, based on characteristics of cell morphological changes and DNA laddering. Using TUNEL staining we observed the DNA fragmentation in SF-21 cells, and further found that the level of apoptosis induced by N.
rileyi cultured fluid was a dose-dependent response. The induction of apoptosis in SF-21 cells was inhibited by adding pan-caspase inhibitor, z-VAD-fmk, indicating that the apoptosis was mediated by caspase. We detected that cleavage signal of DEVD, a sf-caspase-1 substrate peptide,
raised in SF-21 cells after treatment with N. rileyi cultured fluid, and that the apoptosis could be inhibited by adding sf-caspase-1 inhibitor z-DEVD-fmk. These results indicate that sf-caspase-1 is involved in the apoptosis induced by N. rileyi. Furthermore, apoptosis in SF-21 cells
could be inhibited greater by adding both caspase-8 inhibitor, z-IETD-fmk, and caspase-9 inhibitor, z-LEHD-fmk, than adding either one of them, suggesting that two caspase pathways could occur in the apoptosis of SF-21 cells induced by N. rileyi. We also found that the cleavage signal of LEHD, a caspase-9 substrate peptide, elevated after treatment with N. rileyi cultured fluid, suggesting that the activity of initiator caspase in SF-21 cells is similar to that of mammalian caspase-9.
URI: http://hdl.handle.net/11455/30690
其他識別: U0005-0908200711115300
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