臺灣博碩士論文加值系統

近年來雪蓮(Saussurea involucrate)對抗風濕、鎮痛、強心、抗癌、降壓、平喘、清除自由基及抗疲勞等藥理研究取得了可喜進展，其成分主要含多羥基黃酮類化合物，具有明顯的抗炎作用。本研究以雪蓮為研究題材，探討雪蓮是否具有抑制癌細胞生長的能力。以不同乳癌細胞株MCF-7、MDA-MB-453 及正常人類乳房表皮細胞株HBL-100進行細胞生長抑制實驗，結果顯示在MCF-7、MDA-MB 453細胞株中對雪蓮的感受性較好，在100μg/ml濃度下有抑制近50％生長率的效果，顯示雪蓮對於乳癌細胞有較明顯的抑制生長效果。以流式細胞儀檢測雪蓮對MCF-7細胞株細胞週期的影響，結果發現雪蓮會使MCF-7細胞株之細胞週期停滯於G0/G1期。此外，我們更進一步利用西方墨點法測量P21、P27、P53、cyclin D1、CDK4、 p-Erk 1、p-Erk 2及p-JNK這些與細胞週期中G0/G1期相關蛋白質表現情形，結果發現P21、P27及P53在乳癌細胞處理100μg/ml雪蓮後，均有上升的趨勢，CDK4及p-JNK沒有變化，而cyclin D1及p-Erk 1、p-Erk 2則有下降情形。綜合以上實驗結果顯示，經雪蓮作用後， P21、P27及P53表現量上升，導致乳癌細胞分裂停留在G0/G1時期。

Saussurea involucrate is a well-known Chinese herbal which holds many therapeutic effects. Folk applications include easing rheumatoid arthritis, free-radical clearance, anti-tumor and anti-hypertension. Saussurea involucrate, containing bunch of alkaloids and flavonoids, was also used as an anti-inflammatory drug in Chinese medical application. In this study, we focus on the anti-tumor effect of Saussurea involucrate. We compare the growth inhibition of aggressive breast cancer cell line MCF-7, MDA-MB-453 and normal breast epithelial cell HBL-100. Results showed that both MCF-7 and MDA-MB-453 are more sensitive than HBL-100 to the extract of Saussurea involucrate treatments, for IC50 being 100μg/ml. We also examine the cell cycle alternation by flow cytometry. We found that the extract of Saussurea involucrate could arrest MCF-7 cell cycle on the G0/G1 phase, therefore block its proliferation. Furthermore, we used western blot to assay the concentration discrepancy of G0/G1 phase-associate proteins P21, P27, P53, cyclin D1, CDK4, p-Erk 1, p-Erk 2 and p-JNK. We found that P21, P27, P53 have significant elevation during 100μM Saussurea involucrate treatment, while CDK4 and p-JNK remain constant and cyclin D1, p-Erk 1, p-Erk 2 are decreased during the treatment. In conclusion, the active ingredients of Saussurea involucrate could block breast cancer cell growth by stopping cell cycle progression.

中文摘要…………………………………………………i
英文摘要…………………………………………………ii
目次………………………………………………………iii
圖目次……………………………………………………vi
符號與縮寫………………………………………………vii
第一章 緒論…………………………………………… 1
1.1 雪蓮之簡介
1.1.1 植物形態 1
1.1.2 入藥品種 1
1.2 雪蓮之成分與藥理作用之研究 2
1.2.1 化學成分 2
1.2.2 藥理作用 2
(1) 抗炎鎮痛作用
(2) 清除自由基、抗氧化力、抗疲勞和提高免疫作用
(3) 抗癌作用
1.3 細胞週期之介紹 3
1.3.1 細胞週期的調控 5
1.4 乳癌 6
1.4.1 乳癌成因 7
1.4.2 目前乳癌的治療方法 7
1.5 研究動機 9
1.6 HBL-100、MDA-MB-453及MCF-7 三種細胞介紹 9
第二章 材料與方法…………………………………….11
2.1 雪蓮樣品處理 11
2.2 細胞處理 11
2.3 MTT assay 11
2.4 以流式細胞儀測定細胞週期 12
2.5 以西方墨點法測定蛋白的表現量 13
2.6 利用倒立式位像差顯微鏡檢測檢測雪蓮萃取物對 HBL-100、MDA-MB-453及 MCF-7細胞株細胞型態的影響 14
2.7 免疫螢光染色法 15
第三章 結果…………………………………………… 16
3.1 雪蓮對各種不同乳癌細胞株於不同時間點的影響
3.1.1 雪蓮對HBL-100、MDA-MB-453及MCF-7細胞株在24小時細胞抑制之結果
3.1.2 雪蓮對HBL-100、MDA-MB-453及MCF-7細胞株在48小時細胞抑制之結果
3.1.3 雪蓮對HBL-100、MDA-MB-453及MCF-7細胞株在72小時細胞抑制之結果
3.2 以流式細胞儀檢測雪蓮對MCF-7細胞株細胞週期分佈的影響
3.3 以西方墨點法測量P21、P27、P53、cyclin D1、 p-Erk 1、 p-Erk 2、p-JNK及CDK4蛋白質表現情形
3.4 利用倒立式位像差顯微鏡檢測雪蓮萃取物對 HBL-100、MDA-MB-453及 MCF-7細胞株細胞型態的影響
3.5 利用免疫螢光染色法檢測 MCF-7細胞株受雪蓮處理後，p21的表現量
第四章 討論…………………………………………………20
第五章 參考文獻……………………………………………23
第六章 附圖…………………………………………………27

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