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研究生:楊長霖
研究生(外文):Chang-Lin Yang
論文名稱:藉 HeLa 細胞增加唾液酸酶的濃度可能影響 NK 細胞的毒性能力
論文名稱(外文):Increased Concentration of Sialidases by HeLa Cells Might Influence The Cytotoxic Ability of NK Cells
指導教授:許垤棋
指導教授(外文):Dey-Chyi Sheu
口試委員:許垤棋
口試委員(外文):Dey-Chyi Sheu
口試日期:2014-07-30
學位類別:碩士
校院名稱:大同大學
系所名稱:生物工程學系(所)
學門:工程學門
學類:生醫工程學類
論文種類:學術論文
論文出版年:2014
畢業學年度:102
語文別:中文
論文頁數:56
中文關鍵詞:神經胺酸苷酶神經胺酸苷酶抑制劑唾液酸酶細胞毒性子宮頸癌細胞自然殺手細胞
外文關鍵詞:cervical cancer cell linenatural killer cellneuraminidase inhibitorneuraminidasecytotoxicitysialidase
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根據報導指出癌症有能力去逃離免疫系統,此免疫系統主要來自於自然殺手細胞(Natural killer cell;NK cell),雖然真實機制為複雜的,但是有些抑制者來自癌症細胞所分泌,這些抑制者在此機制中為扮演重要角色。此篇研究的目的在於證明由癌症細胞(HeLa)潛在調控者被釋放出能促成減少NK 細胞的細胞毒性。NK-HeLa 共同培養系統使用測試假設從癌症細胞潛在調控者的存在能促成減少NK 細胞的細胞毒性,當共同培養培養基及培養培養基包含商品化唾液酸酶使用於培養NK細胞,使得NK 細胞的細胞毒性也被抑制,然而,細胞毒性部分由唾液酸酶抑制劑(DANA),HeLa 細胞與 NK 細胞之後的西方墨點法證明在 HeLa 細胞內能增加 Neu2 和 Neu3 表現。實驗結果發現 Neu2 和Neu3 表現在癌症細胞內能牽涉NK 細胞的減少反應,而由唾液酸酶抑制劑修復,提供一新的概念應用於未來癌症處理治療中。
Cancer cells reportedly have the ability to escape from the immunesystem, mainly from natural killer(NK)cells. Although the real mechanismsare complicated, some inhibitors that are secreted from the cancer cellsmight play an important role. This study’s aim was to investigate thepotential mediator released by cancer cells(HeLa)that contributes to thedecreased cytotoxicity of NK cells. When the coculture medium and culturemedium containing commercialized sialidase were used to culture NK cells,the cytotoxicity of the NK cells was also inhibited. However, cytotoxicitywas partially restored by a sialidase inhibitor(DANA).Western blot analysisof the HeLa cells after coculturing with NK cells demonstrated increasedNeu2 and Neu3 expression in HeLa cells. The finding that Neu2 and Neu3expression in cancer cells might be involved in the impaired function of NKcells, which could be restored by a sialidase inhibitor, provides a newconcept that could be applied to the management of cancer.
謝誌 i
摘要 ii
Abstract iii
目次 iv
圖次 vi
第一章 導讀 1
1.1 前言 1
1.2 研究目的 2
第二章 文獻回顧 3
2.1 自然殺手細胞 (NK 細胞) 3
2.2 子宮頸癌 (cervical cancer) 6
2.3 唾液酸酶 (Sialidase) 8
2.3.1 生理作用 9
2.3.2 人類唾液酸酶的四種類型作用10
第三章 材料與方法 15
3.1 實驗儀器 15
3.2 實驗材料 16
v
3.2.1 實驗細胞株16
3.2.2 材料來源 17
3.3 實驗方法 19
3.3.1 NK 與HeLa 細胞之間互相作用 19
3.3.2 細胞毒性試驗 .. 19
3.3.3 唾液酸酶 (Sialidase) 及DANA 處理 20
3.3.4 HeLa 細胞在共同培養系統內唾液酸酶活性及表現之檢測 20
3.3.5 免疫轉印 (Immunoblotting) 21
3.3.6 統計分析 21
第四章 實驗結果 22
4.1 NK 細胞與 HeLa 細胞共同培養後 NK 細胞細胞毒性會減少 22
4.2 增加共同培養之培養液的唾液酸酶活性 24
4.3 唾液酸酶及 DANA 在 NK 細胞毒殺內作用 26
4.4 經由 DANA 修復在 NK-HeLa 共同培養系統內的 NK 細胞之細胞毒性 28
4.5 HeLa 細胞在與 NK 細胞共同培養後的唾液酸酶活性 31
4.6 HeLa 細胞在共同培養內增加 Neu2 及 Neu3 表現 33
第五章 結論與討論36
第六章 參考文獻 41
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