穀胱甘肽轉移酶（Glutathione S-transferase﹔ GST）的主要功能包含維持細胞內氧化還原狀態的平衡，但也可對細胞行許多內源性及外源性毒性物質進行解毒，因此在細胞生理上起著重要的作用。本研究的主要目的在於探討SjGST 與morin 對人類乳癌細胞生長的影響及遷移之機制。先利用SjGST 蛋白進行體外細胞增生實驗，實驗表明，濃度5、10、50、100、200、500、700、1000nM 能促進乳癌細胞株MDA - MB435s 增生，但隨著時間越長低濃度的SjGST 對癌細胞的生長越明顯。在轉移試驗方面，低濃度的SjGST 可在第36 與48 小時，
明顯促進癌細胞遷移。另外，在眾多文獻中顯示， Morin（桑色素）具有抗氧化、抗自由基及抑制癌細胞生長的作用。因此，本研究更有興趣知道SjGST 和morin 與癌細胞共處時，SjGST 是否能幫助癌細胞脫離morin 的毒殺反應，以運用於未來癌症藥物治療。但經脫毒實驗
結果顯示，無論是先SjGST、後加SjGST 或是將混合好的SjGST 與morin 一起加入與乳癌細胞MDA-MB435s 作用，都無法幫助細胞脫離morin 的毒殺。

The major functions of glutathione S-transferase (Glutathione S-transferase; GST) include the maintenance of intracellular redox state of substances, and detoxification of cells of endogenous and exogenous toxic substances, and therefore GST plays an important physiological role.
The main purpose of this study is to find the effects of SjGST and morin on human breast cancer cell on growth and migration , and the possible mechanisms. Therefore we first carried out cell proliferation assays in vitro . Results showed that at low concentrations of SjGST could enhance the proliferation of breast cancer cell line MDA-MB435s better then high concentrations.And the cell migration assays results showen, when 36 and 48 hours that the low concentration SjGST can enhance the breast cancer cell line MDA-MB435s migration. Morin have antioxidant,anti-free radicals, and inhibitor of cancer cells growth. Therefore, we aimed at revealing the effect of more, GST and the mixture of both on growth and detoxination of MDA-MB435s. However, the results of detoxification assays shown,even add SjGST first or last or both mixed ,the concentration 5、10、50、100、200、500nM of SjGST still not protect the breast cancer cells (MDA-MB-435s cells line) to break away from morin .

目錄
圖目錄------------------------------------------------------------------------------I
縮寫------------------------------------------------------------------------------II
中文摘要-------------------------------------------------------------------------IV
英文摘要--------------------------------------------------------------------------V
第一章 緒論----------------------------------------------------------------------1
1.1 榖胱甘肽轉移酶（Glutathione S-transferase; GST）--------1
1.2 穀胱甘肽（Glutathione; GSH）--------------------------------2
1.3 榖胱甘肽轉移酶之解毒代謝過程--------------------------------------4
1.4 氧壓與有毒物質之代謝--------------------------------------------------6
1.5 SjGST 的結構------------------------------------------------------------7
1.6 桑色素(Morin)的構型與特性-------------------------------------------8
1.7 MDA-MB435s 細胞株--------------------------------------------10
1.8 研究動機----------------------------------------------------------------11
第二章 材料與方法------------------------------------------------------------12
2.1 實驗材料-----------------------------------------------------------------12
2.1.1 E.coli 菌株-----------------------------------------------------------12
2.1.2 細胞株---------------------------------------------------------------12
2.1.3 實驗儀器------------------------------------------------------------12
2.1.4 實驗藥品-----------------------------------------------------------13
2.1.5 實驗溶液配製-----------------------------------------------------14
2.2 實驗方法--------------------------------------------------------------15
2.2.1 SjGST 蛋白的純化----------------------------------------------15
2.2.2 SDS-PAGE--------------------------------------------------------16
2.2.3 SjGST 蛋白液的透析及濃度測定----------------------------17
2.2.4 細胞培養----------------------------------------------------------18
2.2.5 細胞增生試驗-----------------------------------------------------19
2.2.6 桑色素IC50 濃度測試------------------------------------------20
2.2.7 SjGST 脫毒實驗-------------------------------------------------21
2.2.8 細胞遷移分析---------------------------------------------------21
2.2.9 統計分析-----------------------------------------------------------22
第三章 結果---------------------------------------------------------------------23
3.1 SjGST 蛋白的純化---------------------------------------------------23
3.2 SjGST 增生試驗-------------------------------------------------------23
3.3 Morin（桑色素）IC50 試驗-------------------------------------25
3.4 SjGST 脫毒試驗-------------------------------------------------------26
3.5 SjGST 與細胞遷移試驗----------------------------------------------27
第四章 討論--------------------------------------------------------------------28
4.1 SjGST 蛋白對乳癌細胞增生與遷移影響-------------------------28
4.2 SjGST 對morin 脫毒之影響----------------------------------------29
參考文獻--------------------------------------------------------------------------32
圖表---------------------------------------------------------------------------------------38
圖目錄
圖一、透析後SDS-PAGE電泳圖---------------------------------------------38
圖二、細胞增生試驗------------------------------------------------------------39
圖三、Morin（桑色素）IC50 試驗--------------------------------------------40
圖四、Morin（桑色素）IC50 試驗圖片-----------------------------------41
圖五、SjGST 脫毒試驗一-----------------------------------------------------44
圖六、SjGST 脫毒試驗二-----------------------------------------------------45
圖七、SjGST 脫毒試驗三-----------------------------------------------------46
圖八、SjGST 遷移試驗--------------------------------------------------------47
附圖一、哺乳類動物之榖胱甘肽轉移酶(Glutathione S-transferase﹔
GST)五大類型-------------------------------------------------------------------48
附圖二、肝細胞解毒步驟------------------------------------------------------49
附圖三、榖胱甘肽的氧化與還原結構式------------------------------------50
附圖四、苯(a)芘解毒代謝圖---------------------------------------------------51
附圖五、Phase II（conjugation pathway）接合性途徑-------------------52
附圖六、SjGST 三級結構-----------------------------------------------------53
附圖七、Morin 化學構型------------------------------------------------------54
附圖八、ATCC MDA-MB435s 顯微鏡下型態-----------------------------55

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