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  • 學位論文

墨旱蓮萃取物抑制人類口腔癌細胞轉移及侵襲之機制探討

Study of the inhibitory effects of Eclipta prostrata extract on the migration and invasion of oral cancer cells

指導教授 : 楊順發

摘要


惡性腫瘤在最近幾年一直位居國人十大死因的第一名,其中口腔癌(Oral cancer),又位居十大死亡癌症中的第五名,其嚴重性不可輕忽。而癌症會造成病患死亡主要的原因,是因為其具有遠端轉移(Metastasis)的特性,因此阻斷癌細胞移動的能力,是癌轉移研究的主要目標。癌轉移是個複雜的過程,需要多種因子參與調控,近年來在許多研究中證實了Matrix metalloproteinases (MMPs)參與調控癌細胞的轉移。本篇研究所使用的中草藥墨旱蓮萃取物(Eclipta prostrate extract, EPE),在以往文獻中發現具有抑制肝癌、乳癌及肺癌癌細胞轉移和血管新生(Angiogenesis)的能力,但是對於口腔癌的影響並不清楚。於是本研究使用口腔癌細胞株SCC9來探討,墨旱蓮是否能抑制SCC9細胞株的轉移,以及對MMP-2具有何種影響。首先以MTT assay,得知墨旱蓮萃取物濃度0-100 μg/ml的劑量下,不影響SCC9細胞株細胞存活率,接著以Wound healing及Boyden chamber assay觀察墨旱蓮是否會影響SCC9的移動能力,結果發現,隨著藥物濃度的增加,細胞的移動能力逐漸被抑制,再來以Gelatin zymography及Western blot觀察墨旱蓮對口腔癌細胞株SCC9的MMP-2活性和蛋白表現有何影響,從結果觀察到,墨旱蓮確實會抑制MMP-2的酵素活性及蛋白表現;接著,觀察墨旱蓮對訊息傳遞路徑的影響,發現墨旱蓮抑制了ERK的磷酸化表現;最後利用ERK抑制劑確認墨旱蓮能透過訊息傳遞路徑ERK來調控下游蛋白表現,從以上實驗觀察到的現象,推論墨旱蓮可經由抑制ERK的磷酸化,降低MMP-2的表現,達到抑制口腔癌細胞SCC9的移動能力。

關鍵字

墨旱蓮 口腔癌 轉移 侵襲

並列摘要


Oral cancer has the fifth highest incidence among the different forms of cancer. Metastasis involves multiple sequential cascades including degradation of basement membrane and extracellular matrix (ECM) by the matrix metalloproteinases (MMPs). Recently reports indicate that Eclipta prostrate extract (EPE) have anti-metastatic ability in the liver cancer, breast cancer and lung cancer cells. However, its effects and mechanism on invasion and metastasis of oral cancer cells are unclear. The present study aimed to examine the antimetastatic properties of EPE in SCC-9 oral cancer cell lines. The results of wound healing assays and boyden chamber assays revealed that EPE significantly reduced the migration and invasion of SCC-9 cells. Gelatin zymography and western blotting analyses showed that EPE significantly inhibited the activity and expression of MMP-2. The results also showed that EPE significantly decreased phosphorylated Erk1/2 levels. Moreover, blocking of the Erk1/2 pathway also enhanced the anti-metastatic ability of EPE in oral cancer cells. In conclusion, EPE inhibited MMP-2 expression and oral cancer cell metastasis by downregulating Erk1/2 signals pathways and, thus, exerts beneficial effects in chemoprevention.

並列關鍵字

Eclipta prostrata oral cancer migration invasion

參考文獻


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