- 學位論文
荷葉黃酮萃取物抑制肥胖因子促進人類乳癌細胞的轉移之作用
The flavonoid-rich extract from Nelumbo Nucifera leave inhibits obesity factor induced breast cancer metastasis
摘要
台灣婦女乳腺癌是最常見的癌症，乳腺癌死亡率佔所有癌症死亡 人數的 28.4％，並且與肥胖有關。先前的研究利用荷葉萃取物(NLE) 已被証實可以有效地減少脂肪肝、高脂血症、動脈粥樣硬化和抗氧化 能力。本研究中利用脂肪細胞分泌肥胖因子而得的培養液(CM)誘導 人類乳癌細胞遷移的模式，並以 Boyden chamber 驗證，從結果得知當給予 NLE 時可以減緩在 CM 誘導狀態下乳腺癌細胞的遷移作用，隨著 NLE 劑量增加抑制的效果更明顯，且我們亦使用 ECIS 及傷口癒合試驗加以確認，最後所得到的結果皆一致。為了探討乳癌細胞遷移的分子機制，利用二維膠體電泳分析，從結果中得知在 CM 誘導的狀態下，細胞 Hsp90、StarD10、Peroxiredoxin 6 蛋白表現較高，推測NLE或許是透過這些蛋白進而影響遷移的能力；再者，我們使用西方墨點法分析人類乳癌細胞株(MCF-7)在 CM 誘導的狀態下，並且處理NLE觀察其蛋白的表現，從結果得知當處理NLE時會降低 JNK、PI3K/AKT、StarD10 和 Peroxiredoxin 6 等蛋白表現量，進而達到抑制人類乳癌細胞的遷移作用。綜上所述，本研究的結果表明未來對於治療轉移性乳癌，NLE 具有發展成為抑制癌症轉移過程中的天然藥物之潛力。
並列摘要
Breast cancer is the most common cancer in women in the Taiwan. Then breast cancer deaths accounted for 28.4% of all the cancer deaths, and have been associated with obesity. Previous studies, has been confirmed Nelumbo nucifera leave extract (NLE) can effectively reduce the fatty liver, atherosclerosis, hyperlipidemia and anti-oxidative properties. Thus, we investigated the relationship between obesity and breast cancer, and evaluated the inhibitory effect of NLE on the adipocytes-induced breast cancer progression. Adipocyte secreted conditioned medium (CM) noticeably induced breast cancer migration. NLE diminished the CM-induced migration number of breast cancer cell by boyden chamber assay. A direct correlation was found between adipocyte and breast cancer cell migration. Similarly, we used of ECIS assay, wound healing assay and transwell assay identical results were found. To investigate the molecular mechanisms of breast cancer cell migration using two-dimensional gel electrophoresis. We found CM increased the proteins level of the Hsp90, StarD10, Peroxiredoxin 6. The results suggest that CM stimulates migration of breast cancer cells, perhaps through the interaction of these proteins. Under the CM treatment, phosphorylated JNK, PI3K/Akt, StarD10 and Peroxiredoxin 6 levels of MCF-7 cells treated with NLE can significantly inhibited the protein expression were measured by Western blot analysis. These data indicate adipocytes provide a major component of the microenvironment for rapid tumor growth and migration, and this increase was inhibited significantly by NLE treatment. In summary, the study indicated that the future for the treatment of metastatic breast cancer, NLE inhibited cancer metastasis has developed into a process of natural drugs.