- 學位論文
蘆丁抑制攝護腺癌細胞LNCaP之效果探討
Anti-Cancer Effect of Rutin in Human Prostate Cancer Cell LNCaP
摘要
從先前的研究，發現蘆丁 (Rutin) 藉由調節各種標靶分子，包括TNF-α 及 c-met ，有抗癌的特性，此研究在於探討蘆丁對於攝護腺癌的調控。 我們使用兩種細胞株 (LNCaP 及LNCaP Enz-R) ，而 LNCaP Enz-R 是用5 μM 的 Enzalutamide 治療六個月後， 所篩選出來對於Enzalutamide 有著抗藥性的細胞株。我們使用四種不同濃度的蘆丁藥劑(0 μM、25 μM、50 μM 及100 μM) ，經過24 小時，我們利用細胞存活率分析來測定細胞的活性；利用細胞遷移分析來測定癌細胞遷移能力；藉由西方墨點法來測定攝護腺癌細胞進展時的活性氧製造、粒線體生合成及粒線體內標靶分子；免疫螢光染色可用以偵測上皮細胞間質轉換 (EMT) 及其相關的因子。蘆丁顯著抑制了 LNCap 和 LNCap Enz-R 細胞增殖和遷移。 此外，蘆丁增強了 LNCap 和 LNCap Enz-R 細胞中超氧化物歧化酶 (SOD) 活性催化的調節。我們也發現蘆丁上調控了前列腺癌細胞粒腺體生合成的表現。我們證明了蘆丁透過抑制 EMT 標記(Snail 和 Slug) 與增強 E-cadherin 的表現，來抑制攝護腺癌。更重要的是，蘆丁顯著增加了粒腺體因子的表現，進而誘導 AMP activatedprotein kinase (AMPK) 激活。我們的研究結果表明，對於攝護腺癌來說，蘆丁可當作一種輔助治療方法。
關鍵字
無並列摘要
Rutin has been demonstrated their antitumor activity by modulating various macromolecular targets. This study will explore the regulation of rutin to the prostate cancer. LNCap was treatment with 5μM enzalutamide for more than 6 months and this enzalutamide-resistance of LNCaP cells designated as LNCap Enz-R. The LNCap and LNCap Enz-R were treated with various concentrations of rutin (0, 25, 50, 100 μM) for 24 hours followed by the examination of cell viability using MTT and trypan blue exclusion assays. Cell migration was assessed by wound healing assay. We investigated the ROS production, mitochondrial biogenesis and mitochondrial marker (e.q. AMPK) during the progression of prostate cancer by kit and Western blotting assay. Western blot and immunofluorescence staining were used to detect the expression of epithelialmesenchymal transition (EMT)-associated factors, EMT signaling and AMPK axis. Rutin treatment significantly inhibited cell proliferation and migration of both LNCap and LNCap-Enz-R cells. In addition, rutin enhanced SOD activity-mediated modifications in both LNCap and LNCap-Enz-R cells. Also, we showed that the expression of mitochondrial biogenesis of prostate cells was upregulated by rutin. We demonstrated that rutin exhibited the anticancer effects on prostate cancer via inhibition of expression of EMT marker, Snail and Slug, with increase expression of E-cadherin. More importantly, treatment with rutin significantly increased the expression of mitochondrial factor subsequent to induced AMPK activation. Collectively, our findings revealed that rutin is a promising adjunctive therapy for prostate cancer treatment. These findings indicate that rutin can be effectively inhibited prostate cancer cell by means of various macromolecular targets.