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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/17614

Title: 三氧化二砷誘發鰻魚鰓細胞產生差異性蛋白質之探討
Differential protein expression profiles in Anguilla anguilla eel cell exposed to Arsenic Trioxide (As2O3)
Authors: Hui-Chen Chu
朱惠真
Contributors: NTOU:Institute of Bioscience and Biotechnology
國立臺灣海洋大學:生物科技研究所
Keywords: ;細胞;蛋白質;二維電泳
arsenic;cell line;protein;2-D
Date: 2002
Issue Date: 2011-07-04
Abstract: 砷在自然界中會以有機砷及無機砷的形式存,其中以無機砷的毒性較強。在水域環境中,魚體因為用鰓來交換氣體,這時會在鰓組織上產生離子的交換,砷此時也會因為離子交換的過程而有可能進入到生物體內。 使用7.5μM的As2O3作用鰻魚34代細胞 (peimary cell 34 passage) 時,細胞在型態上並沒有顯著的改變,但是在可溶性蛋白已有顯著的變化,以Sodium dodecyl sulfate- polyacrylamine gradient electrophoresis(SDS-PAGE)及Two dimension electrophoresis(2-D)分析,發現砷在作用一小時、三小時及六小時之後,分子量為60kDa、40kDa及27kDa蛋白質處有顯著的差異出現,而該群蛋白質等電點聚焦在pI 5-6處。我們從第34代鰻魚鰓細胞選殖AJ2034細胞進行大量培養後,以7.5μM的As2O3處理,並以Cu2+為正對照,在作用40分鐘後,由SDS-PAGE及2-D上發現,出現在分子量為70kDa處有單一的蛋白質產生。因此我們推估,在低等脊椎動物細胞上,當細胞尚未有明顯外觀上的傷害時,在短時間之內As2O3所刺激鰻魚鰓細胞所產生的90kDa、70kDa、60kDa、40kDa及27kDa的蛋白質當之中,70kDa是最早表現出來的,且AJ2034細胞為主要的反應細胞,同時此蛋白質也可作為鰻魚細胞對砷刺激後具有專一性的Biomarker。
Arsenic is a metalloid with two biologically important oxidation states , AsⅤand AsⅢ. In the nature environment, arsenic exist in both forms as organic and inorganic compounds .In general, inorganic arsenic compound are more toxic than organic arsenic compounds. In water environment, fish exchange the gas with gill, and the iAs enter the living organism through this passway. When we stimulate 34 passage eel gill cell to induce protein by 7.5μM As2O3, we find the morphology dose not change a lot. Nevertheless, soluble protein changes a lot from the results of SDS-PAGE and Two Dimensional Electrophoresis, and we find there have clearly difference in the protein molecular weight of 60kDa,40kDa and 27kDa when stimulated cells within six hours, those protein are focus pI on 5 to 6. Once the AJ2034 cells was cloned and cultured with cells from 34th passage eel gill cell, and when those cells were stimulated by 7.5μM As2O3 with a equal concentration copper as positive control for 40 minutes, we obtained a single 70kDa protein expression results either by SDS-PAGE or 2-DE, Therefore, although the low level vertebrate has not appeared distinct damage in the cell morphology, for those proteins of 90、70、60、40 and 27kDa extracted from eel gill cells which stimulated by As2O3 for 40 minutes, the 70kDa protein was expressed in advance and AJ2034 is the particular expression cell, and 70kDa protein could be considered as a biomarker when eel gill cells stimulated by As2O3.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#GAAAAAAAAAC
http://ntour.ntou.edu.tw/ir/handle/987654321/17614
Appears in Collections:[生命科學暨生物科技學系] 博碩士論文

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