臺灣博碩士論文加值系統

中文摘要
在最近幾年的研究中，癌細胞的醣解代謝已經變成癌症重要特徵之一，因為有氧醣解 (aerobic glycolysis) 會提供癌細胞生長與分裂所需要的能量和材料。並且在許多文獻中提出Pyruvate kinase M2 (PKM2) 可以促進癌細胞的醣解代謝走向有氧醣解(1-3)，Warburg effect就是一個有名的例子。而我們在一篇論文中看到Shikonin，由紫草根萃取出來的萘醌化合物，可以透過抑制腫瘤性PKM2的活性進而有效地抑制乳癌細胞的生長。因此在這篇論文我們測試了Shikonin是否能抑制卵巢癌細胞的生長。根據我們的實驗結果，顯示Shikonin可以有效降低卵巢癌細胞生長速率，並且導致卵巢癌細胞的細胞走向細胞凋亡。此外我們以醣利用率和乳酸生成率分析驗證了Shikonin確實可以干擾卵巢癌細胞的有氧醣解。更進一步地我們也發現到Shikonin可以有效地抑制癌細胞的爬行和貼覆能力。在臨床上，卵巢癌病患的腫瘤大多數為惡性，而從我們實驗的結果顯示PKM2表現在惡性腫瘤明顯的高於良性的。綜合本篇論文的結果我們發現到Shikonin可以有效的抑制PKM2活性及卵巢癌細胞的生長，在治療卵巢癌上可能具有很大的潛力，並且有機會作為對一般化療無效病患的第二線選擇性用藥。

Abstract
Recently, special taxis of glucose metabolism have been proved as an important hallmark of cancer, because glycolysis provides fuel for cell growth and division in cancer cells. Many studies demonstrated that pyruvate kinase (PK)-M2 (PKM2) could transit glucose metabolic flux to aerobic glycolysis as known as the Warburg effect in cancer cells. Shikonin, a natural naphthoquinone isolated from the traditional Chinese herb, has been reported to inhibit tumor-specific PKM2 activity and to result suppression of breast cancer cells proliferation.
In this study, we attempted to clarify that whether shikonin would inhibit the cell growth or induce cell death in several ovarian cancer cell lines. We found that shikonin could induce cell apoptosis and repress ovarian cancer cells proliferation. Furthermore, we confirmed that decrease of glucose consumption and lactate production in shikonin-treated ovarian cancer cells, and we speculated the effect of shikonin is through inhibition of PKM2 activity. In addition, we also found that shikonin inhibited migration and adhesion capacities of ovarian cancer cells. The majority tumors of ovarian cancer usually are malignant. In our data, the statistics of clinical samples in ovarian cancer patients show the positive relationship between PKM2 and malignant tumor.
Together, we conclude that shikonin showed a therapeutic potency for ovarian cancer treatment. It might be as one of second-line drugs for ovarian cancer therapeutics，because shikonin have the ability to repress PKM2 activity of ovarian cancer cells.

中文摘要．．．．．．．．．．．．．．．．．．．．．．．．．．．．．4
英文摘要．．．．．．．．．．．．．．．．．．．．．．．．．．．． 5
第一章 緒論．．．．．．．．．．．．．．．．．．．．．．．． 7
第一節 卵巢癌的簡介．．．．．．．．．．．．．．．．．．．．． 8
第二節 PKM2基因的簡介．．．．．．．．．．．．．．．．．．． 11
第三節 Shikonin的簡介．．．．．．．．．．．．．．．．．．．． 12
第二章 實驗目的與實驗設計．．．．．．．．．．．．．．．．．．．． 13
第三章 實驗材料與方法
第一節 實驗材料．．．．．．．．．．．．．．．．．．．．．．． 14
第二節 實驗方法．．．．．．．．．．．．．．．．．．．．．．． 15
第四章 實驗結果
第一節 Shikonin 可以減緩卵巢癌細胞的生長．．．．．．．．．．． 18
第二節 Shikonin 可抑制卵巢癌細胞代謝和惡性特徵．．．．．．．． 19
第三節 PKM2表現在卵巢癌的臨床意義．．．．．．．．．．．． ． 21
第五章 討論．．．．．．．．．．．．．．．．．．．．．．．．．． 22
第六章 結論與展望．．．．．．．．．．．．．．．．．．．．．．．． 24
參考文獻．．．．．．．．．．．．．．．．．．．．．．．．． ．．． 25

圖表目錄
圖一、PKM2 mRNA 在卵巢癌細胞中的表現．．．．．．．．．．．．．． 27
圖二、Shikonin對卵巢癌細胞生長速率的影響．．．．．．．．．．．．． 28
圖三、Shikonin對卵巢癌細胞細胞週期的影響．．．．．．．．．．．．． 29
圖四、Shikonin對卵巢癌細胞 ATP 生成的影響．．．．．．．．．．．．30
圖五、Shikonin對卵巢癌細胞的醣代謝利用率之影響．．．．．．．．．． 31
圖六、Shikonin對卵巢癌細胞的乳酸生成率之影響．．．．．．．．．．． 32
圖七、Shikonin對卵巢癌細胞爬行能力的影響．．．．．．．．．．．．． 33
圖八、Shikonin對卵巢癌細胞貼附能力的影響．．．．．．．．．．．．． 34
圖九、PKM2在卵巢癌臨床檢體中的表現．．．．．．．．．．．．．．． 35

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