臺灣博碩士論文加值系統

肺癌是近年來臺灣地區致死率最高的癌症，不同種類的肺癌當中又以肺腺癌的致死率最高。前人研究已指出，山苦瓜能抑制乳癌與前列腺癌細胞株的生長。本論文以不同的山苦瓜乙酸乙酯-乙醇(ethyl acetate - ethyl alcohol, Et-EA)萃物之下游萃物來處理人類肺腺癌細胞株CL1-0與CL1-5，觀察是否會抑制CL1-0與CL1-5細胞的生長(proliferation)與移行(migration)，四種乙酸乙酯-乙醇萃物之下游萃物分別為：純水萃物[萃物A]、50%甲醇萃物[萃物B]、100%丙酮萃物[萃物C]、50%甲醇萃物經酸水解所得之萃物[萃物D]。實驗結果顯示：(1) 高濃度的[萃物C] (500 μg/ml)與[萃物D] (300 μg/ml)明顯降低CL1細胞的增生或造成細胞死亡。這代表在山苦瓜Et-EA的下游萃物當中，[萃物C]與[萃物D]能抑制CL1-0與CL1-5細胞的生長。隨著處理的萃物濃度越高，抑制生長的效果越明顯。另兩種Et-EA下游萃物則無明顯抑制CL1-0與CL1-5細胞生長之效果。(2) 在100 μg/ml 的[萃物C]與[萃物D]的處理下，即可抑制CL1-0與CL1-5細胞的移行，隨著處理的萃物濃度越高，抑制移行的效果越明顯。(3) 200 μg/ml的[萃物C]與[萃物D]能抑制細胞生長，並且能抑制CL1-0與CL1-5細胞的移行相關蛋白ADAM15與CD44表現，但不會造成CL1-0與CL1-5細胞死亡。(4) 比起[萃物C]，[萃物D]對CL1-0與CL1-5細胞的藥效較強。我們的結果顯示：山苦瓜之乙酸乙酯-乙醇之萃物對肺癌細胞株的增生、死亡、移行等均有明顯之影響，因此我們合理推測山苦瓜之乙酸乙酯-乙醇之萃物具有成為抗肺癌藥物的可能性。

The fatality rate of lung cancer is the highest among all kinds of cancer in Taiwan. It has been demonstrated that bitter melon (Momordica charantia) extracts (BMEs) can inhibit the proliferation of breast cancer and prostate cancer cell line. We use four different “ethyl acetate-ethyl alcohol (Et-EA)” BMEs to treat human lung carcinoma cell lines CL1-0 and CL1-5 and investigate the effects of BMEs on these cell lines. The four down-streamed Et-EA BMEs used are pure water extract [BME-A], 50% methanol extract [BME-B], 100% acetone extract [BME-C] and acid-hydrolyzed (AH) products of 50% methanol extract [BME-D]. The results revealed that: (1) 500 μg/ml [BME-C] extract and 300 μg/ml [BME-D] can inhibit the proliferation of CL1-0 and CL1-5 cells, and the higher concentration of [BME-C] or [BME-D], the more significant effect in inhibiting CL1 cells proliferation. Other two Et-EA down-streamed BMEs have no effect to CL1 cells in proliferation. (2) 100 μg/ml [BME-C] and [BME-D] can inhibit the migration ability of CL1-0 and CL1-5cells. The higher concentration of 100% acetone extract or [BME-D] exhibits significant effect to inhibit CL1 cells migration. (3) 200 μg/ml [BME-C] and [BME-D] not only inhibit the proliferation of CL1-0 and CL1-5 cells, but also have no effect to cell death. Under 200 μg/ml [BME-C] or [BME-D] treatment, the expression of migration-associated protein, ADAM and CD44, are inhibited.(4) Compare to [BME-C], [BME-D] can exert stronger effect to CL1-0 and CL1-5 cells in proliferation and migration.
Taking together, we conclude that BMEs can inhibit the proliferation, migration and invasion of the lung cancer lines CL1-0 and CL1-5. It is suggested that the BMEs can be a potential anti-lung cancer therapeutic drug.

目錄
中文摘要.................................................................................................... i
英文摘要................................................................................................... ii
縮寫對照表.............................................................................................. iv
一、緒言(Introduction).............................................................................. 1
1.1 肺癌.......................................................................................... 1
1.2 山苦瓜...................................................................................... 2
二、目的(Purposes)................................................................................... 5
三、材料與方法(Materials and Methods)................................................. 6
3.1 化學藥品(Chemicals)................................................................ 6
3.2 細胞培養(Cell culture).............................................................. 6
3.3 細胞存活率測定(MTT assay).................................................. 7
3.4 Hoechst染色(Hoechst assay)................................................. 7
3.5 傷口復原移行實驗(Wound-healing migration assay)............. 8
3.6 Transwell移行實驗(Transwell migration assay)................... 8
3.7 西方墨點轉漬法(Western blotting).......................................... 8
3.8 統計分析(Statistical analysis)................................................. 10



四、結果(Results)................................................................................... 11
4.1 高濃度的特定山苦瓜萃物降低CL1-5、CL1-0肺腺癌細胞增生或造成細胞死亡..................................................................... 11
4.2 濃度200μg/ml的山苦瓜萃物會抑制CL1-5細胞的生長但不造成其死亡................................................................................. 11
4.3 高濃度的山苦瓜萃物抑制CL1-5細胞的移行能力.............. 12
4.4 高濃度的山苦瓜萃物抑制CL1-5細胞的蛋白質ADAM15與CD44表現.............................................................................. 13
五、討論(Discussion)............................................................................. 14
5.1 山苦瓜萃物降低人類肺腺癌細胞株CL1-0與CL1-5存活度的可能原因................................................................................. 14
5.2 山苦瓜萃物是否會影響人類肺腺癌細胞株CL1-0與CL1-5的轉移能力？............................................................................. 15
5.3 更進一步的山苦瓜下游萃物分析.......................................... 15
5.4 山苦瓜Et-EA的下游萃物對其他癌症細胞株的影響.......... 16
六、結論(Conclusions)與未來擬繼續進行之研究工作(Further work). 17
七、參考文獻 (References)................................................................... 18
八、圖片說明(Figures and Figure Legends).......................................... 21

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