中草藥抑制乳癌細胞侵入能力及誘導細胞凋亡之機制探討

Study of the mechanism of Chinese medicinal herbs on the anti-metastasis and inducing apoptosis of breast cancer cells

詹雅筑(Ya-Chu Chan)

指導教授 ： 謝易修

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2007年)

https://doi.org/10.6834/CSMU.2007.00003

摘要

癌細胞的轉移和抗細胞凋亡都是惡性腫瘤的重要特徵，並且也是 在臨床治療上最難以根治及治療複雜度提昇的主要原因。因此，如何 治療癌症病人、開發新藥或是利用天然藥物觀察是否會抑制癌細胞轉 移是一個迫切的問題。長期以來，中草藥被應用於傳統醫學及民俗療 法中的各種治療，並且已有多種中草藥被確認其具有抗氧化、抗發 炎、抗過敏及抗癌的活性，但是到目前為止，對於中草藥抑制癌細胞 轉移、誘導癌細胞凋亡以及其中之詳細機轉的都還不是很清楚。在本 篇論文中，我們利用補骨脂、欖仁樹的葉、魚腥草與半枝蓮之酒精 (50%)萃取物觀察其抑制多種乳癌細胞(MDA-MB-231、JC 與Hs 578T) 生長及抗轉移的功效，證實補骨脂萃取物(Psorlaea corylifolia L. extracts; PCE)隨著濃度的增加，可抑制乳癌細胞(MDA-MB-231 與JC 細胞)的活細胞數並增加死亡的細胞數目，以DAPI stain 與DNA 斷裂 分析方法，發現補骨脂萃取物會導致chromatin condensation 與DNA 斷裂並進一步促使癌細胞走向凋謝死亡。在動物實驗中，我們利用 BALB/c 老鼠於皮下種植JC 老鼠乳癌細胞，並餵食補骨脂萃取物發 現其會抑制腫瘤在活體的生長。欖仁樹的葉萃取物(Terminalia catappa L extracts; TCE)也可透過chromatin condensation 使JC 細胞走向凋 亡。在抗細胞轉移方面，魚腥草萃取物(Houttuynia cordata extracts; HCE)與半支連萃取物(Scutellaria barbata extracts; SBE)可顯著抑制具 有高度轉移能力之人類乳癌細胞(Hs 578T)的侵入及移動能力，利用 gelatin-及casein-zymography 分析方法，探討細胞外基質降解相關基 因，如matrix metalloproteinase-2 (MMP-2)、urokinase-type plasminogen 2 activator (u-PA)、tissue inhibitor matrix metalloproteinase -2 (TIMP-2) 與plasminogen activator inhibitor-1 (PAI-1)的蛋白表現，結果發現魚腥 草萃取物與半枝蓮萃取物對 u-PA 的分泌量有明顯的抑制，並呈現濃 度的效應關係，此外只有魚腥草萃取物也可明顯抑制細胞TIMP-2， 但對於PAI-1 之蛋白表現則是沒有影響，深入探討其作用機轉，結果 顯示半枝蓮萃取物可顯著抑制 p-ERK1/2 。綜合以上結果，推測補骨 脂萃取物與欖仁樹的葉萃取物可以誘導癌細胞的凋亡；魚腥草萃取物 與半枝蓮萃取物可透過抑制腫瘤細胞株 u-PA 分泌的作用進而抑制 癌瘤細胞遷移和侵犯作用的能力，推論可應用於輔助癌症的治療，以 提昇抗癌藥物療效。

關鍵字

中草藥, 乳癌細胞, 細胞凋亡

並列摘要

Both the metastasis and anti-apoptosis of cancer are a vital trait in
malignance with extreme difficulties in early diagnosis and therapeutic
management. So far, the most important thing for finding new cancer
drug is urgent. Chinese medicinal herbs have been used for therapeutic
purposes in traditional and folk medicine for a long time. A recent report
showed that many kinds of Chinese medicinal herbs have the antioxidant,
anti- inflammation, anti-allergic and anti-tumor activeness. However, the
effects and detailed mechanisms of Chinese medicinal herbs on cancer
cell metastasis and inducing cell apoptosis were still unclear. In this study,
anti-tumor and anti-metastasis agents were investigated using 50%
alcohol extracts of Psorlaea corylifolia L., Terminalia catappa L.,
Houttuynia cordata, and Scutellaria barbata on various breast cancer
cells (MDA-MB-231, Hs 578T, JC). We demonstrated that Psorlaea
corylifolia L. extracts (PCE) induced cell death, chromatin condensation,
and DNA fragmentation on MDA-MB-231 and JC cells. Furthermore,
PCE were evidenced by its inhibition on the growth of JC cells in vivo.
Terminalia catappa L extracts (TCE) also exerted an anti-tumor effect
through an increase of dead cells and chromatin condensation on JC cells.
For anti-metastasis, in the absence of cytotoxicity, Houttuynia cordata
extracts (HCE) and Scutellaria barbata extracts (Pereira,
Strasberg-Rieber et al.) exerted a dose-dependent inhibitory effect on the
invasion, and motility of highly metastatic human breast cancer cells (Hs
4
578T). We examined the effects of HCE and SBE on factors of cancer
metastasis. We treated tumor cells with various concentrations of HCE
and SBE, for set periods, and then subjected cells to gelatin zymography,
casein zymography, and Western blot to investigate the expression of
matrix metalloproteinase-2 (MMP-2), urokinase-type plasminogen
activator (u-PA), tissue inhibitor matrix metalloproteinase -2 (TIMP-2),
and plasminogen activator inhibitor-1 (PAI-1). Following treatment with
these compounds was found to decrease the expression of u-PA in a
concentration-dependent manner and only HCE could reduce the
expression of TIMP-2. Further, a treatment of SBE also resulted in an
inhibition on the activation of p-ERK1/2. Together, these results suggest
that PCE and TCE could induce cell apoptosis; HCE and SBE may act to
decrease the in vitro invasiveness of cancer cells and therefore, reduce the
invasion and metastasis of tumor cells. This study suggests a possible role
for these extracts in cancer therapy.

並列關鍵字

Chinese medicinal herbs, anti-metastasis, apoptosis

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