薑黃素衍生物ASC-J9對MDA-MB-231人類三陰性乳腺癌細胞生長和轉移之影響

Effect of curcumin analog ASC-J9 on cell survival and metastasis in MDA-MB-231 triple-negative human breast cancer cell

王鈺翔(Yu-Shiang Wang)

指導教授 ： 李健群

中山醫學大學/健康管理學院/營養學研究所/碩士(2016年)

摘要

根據衛生福利部民國103年十大癌症死亡統計，女性乳房腫瘤排名第四，相較於其它歐美國家，國內的乳癌女性患者有年輕化趨勢，因此對於如何降低乳癌的發生和致死率，為目前重要研究課題。ASC-J9為薑黃素(curcumin)的衍生物之一，少數研究證實它對前列腺癌具有抑制效果，然而ASC-J9是否對於乳癌具有抗癌效果仍未知。本研究利用細胞和動物模式，探討ASC-J9對高惡性度MDA-MB-231三陰性乳癌細胞株生長、轉移之影響。結果顯示：處理0.1、0.5、1、5 μM ASC-J9 24小時後可顯著抑制MMP-9酵素活性、蛋白質和mRNA表現量並抑制乳癌細胞移行(migration)和侵襲(invasion)。處理ASC-J960分鐘後可顯著抑制JNK和p38磷酸化，若以JNK和p38抑制劑處理細胞亦可降低MMP-9蛋白質表現。以ASC-J9處理MDA-MB-231細胞120分鐘後，可顯著抑制c-jun核蛋白表現量並透過electrophoretic mobility shift assay方法證實ASC-J9可降低c-jun和AP-1 binding site的結合能力。由細胞存活率分析及凋亡檢測實驗結果得知，處理高劑量ASC-J9 (10、20、30 μM ) 24小時後，可改變Bax/Bcl-2的比例並誘發MDA-MB-231細胞caspase 3、PARP蛋白質表現量並促使乳癌細胞壞死(Necrosis )、凋亡(Apoptosis )。由上述結果證明低劑量ASC-J9可透過抑制JNK/p38訊號路徑降低c-jun核內蛋白質累積，進而減少其與AP-1結合能力，抑制MMP-9表現和活性，最終抑制乳癌細胞移行與侵襲。而高劑量ASC-J9則可誘發癌細胞壞死及凋亡。另外以帶有標記冷光酵素(luciferase)的MDA-MB-231三陰性乳癌細胞異體移植免疫缺陷小鼠(C.B17/Icr-Prkdcscid/CrlNarl mouse)為動物模式，餵食ASC-J9 (50 mg/kg)四週後能有效抑制腫瘤生長。綜合上述實驗結果ASC-J9具有抗乳癌細胞發展之潛力。

關鍵字

ASC-J9 ； 基質金屬蛋白酶-9 ； 轉移 ； 凋亡 ； 壞死 ； MDA-MB-231乳癌細胞

並列摘要

Breast cancer is the most prevalent cancer among women worldwide, and it was becoming more common in younger women. Reduction of incidence and death rate is still an important issue in the world. ASC-J9, an analog of curcumin, has been recognized as a potential antioxidant, demonstrated to have anti-cancer effect against human prostate cancer. However, this anti-cancer effect on human breast cancer is not fully clarified. In this study, we investigated the effect of ASC-J9 on cell growth and metastasis on MDA-MB-231 human triple-negative breast cancer cells. Results indicated that ASC-J9 dose-dependently inhibited MMP-9 protein, mRNA expression and enzyme activityas well as cell migration and invasion. The phosphorylation of JNK and P38 were suppressed by ASC-J9 treatment for 60 min. The MMP-9 protein expression were significantly inhibited by specific inhibitors of JNK and p38. Moreover, the nuclear accumulation of c-Jun protein and AP-1 DNA binding activitywere decreased by ASC-J9 treatment for 120 min. Treatment with high concentration of ASC-J9 (10, 20, 30 μM ) were able to change the ratio of Bax/Bcl-2 and induce caspase 3as well as PARP expression and activity, resulted in necrotistic and apoptotic cell death. The MDA-MB-231 breast cancer xenograftmodel in C.B17/ Icr-Prkdcscid/ CrlNarl mouse was used to exam the effect of ASC-J9 on tumor growth. Oral administration of ASC-J9 for four weeks suppressed tumor tumor growth. Taken together, our finding suggests that ASC-J9 has anti-tumor potentials against breast cancer development.

並列關鍵字

ASC-J9 ； Matrix metalloproteinase-9 ； Metastasis ； Apoptosis ； Necrosis ； MDA-MB-231 breast cancer cell

參考文獻

行政院衛生署國健康局(2014):中華民國103癌症登記報告。台北:行政

院衛生署國民健康局編印

行政院衛生署(2014):中華民國103年死因統計結果分析。台北:行政院

衛生署國民健康局編印

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