蛇葡萄素抑制膠質母細胞瘤移行的分子機轉

膠質母細胞瘤 (GBM) 作為最常見的惡性腦腫瘤，具有高度侵襲以及轉移能力，死亡率高且預後不佳，現今的治療手段對於 GBM 效果都有限。中草藥中的黃酮類化合物已被證實對於癌症的治療是有益處的，蛇葡萄素為從蛇葡萄屬植物中萃取出的活性成分，目前已研究出多種功效，例如抗菌、抗氧化，護肝以及抗腫瘤等。本研究旨在探討蛇葡萄素對於膠質母細胞瘤的抗腫瘤功效及其分子機制。透過 MTT assay 顯示處理了 0-200 M 蛇葡萄素的 GBM 8401 與 GBM U251 細胞生長沒有顯著影響。因為毒殺效果不彰，轉而觀察是否能夠抑制 GBM 8401 和 GBM U251 細胞的侵襲轉移能力。利用Boyden chamber 檢測發現隨著藥物濃度上升 GBM 8401 細胞轉移與侵襲的能力明顯的被抑制，但對於 GBM U251 細胞並沒有顯著的效果。Gelatin Zymography 證明了 GBM 8401 細胞中第九型與第二型基質金屬蛋白酶 (Matrix metalloproteinase, MMP-9 and MMP-2) 的活性皆會因藥物濃度提高而下降。最後透過西方墨點法證實了 GBM 8401 中 MMP-9 和 MMP-2 蛋白表現量被蛇葡萄素抑制的效果，磷酸化的細胞外調節蛋白激酶 (phosphorylated extracellular regulatory protein kinase, p-ERK) 表現量也同樣的被抑制。不過上皮間質轉化的標記蛋白 E-cadherin 與 N-cadherin 兩者的蛋白表現量皆沒有什麼改變，但 snail 的活性卻有被抑制。總結上述，確定了蛇葡萄素具有透過降低 MMP-9 與 MMP-2 活性來抑制 GBM 的轉移與侵襲的功效，並認為蛇葡萄素能夠做為治療 GBM 的有效方法之一。

關鍵字

蛇葡萄素；膠質母細胞瘤

並列摘要

Glioblastoma (GBM) as the most common malignant brain tumor has high ability of migration and invasion and caused GBM has poor prognosis. Till now, the treatment of GBM is not effective. Research has shown that flavonoids in traditional Chinese medicines have ability to treat many cancers. Ampelopsin is an effective component extracted from Ampelopsis species and exerts several biological functions such as anti-bacterial, anti-oxidant, liver protection and anti-tumor. The purpose of this study was to investigate the anti-tumor effect and underlying molecular mechanism of ampelopsin on GBM. We found that treatment with 0 to 200 M ampelopsin has no significant effects on cell viability of GBM 8401 and GBM U251 cells. Next, we wanted to investigate the inhibition ability of ampelopsin in migration and invasion of GBM cells. By boyden chamber assay, we found that the ability of migration and invasion were inhibited of GBM cells in proportion to the concentration of ampelopsin. Similarly, the activities of MMP-9 and MMP-2 were inhibited in a dose-dependent manner by ampelopsin as shown by zymography analysis. In addition, western blot analysis revealed that the protein expression of MMP-9 and MMP-2 were decreased after treated with ampelopsin. The level of phosphorylated ERK also decreased in response to ampelopsin treatment. However, the proteins involved in epithelial-mesenchymal transition such as E-cadherin and N-cadherin were not markedly changed, but the expression of snail was inhibited by ampelopsin. Taking all result together, ampelopsin inhibited the migration and invasion of GBM cells by decreased the activity of MMP-9 and MMP-2, and consider that it could be effective agent for treatment of GBM.