探討薑黃素在攝護腺癌細胞移動、侵襲以及間質蛋白酶活化的影響

Suppression of prostate cancer cell migration and invasion by curcumin through down-regulating matriptase activity

林雅筠(Ya-Yun Lin)

指導教授 ： 李明學

國立臺灣大學/醫學院/生物化學暨分子生物學研究所/碩士(2010年)

https://doi.org/10.6342/NTU.2010.02469

摘要

攝護腺癌的癌症致死率在西方男性世界中排名第二位；在台灣，攝護腺癌的發生率以及死亡率亦有逐年提升的現象。攝護腺癌的高致死率主要是由於癌症的病程發展到癌細胞具有較高移動、侵襲和轉移能力或變成雄激素非依賴型的徵狀。薑黃素 (Curcumin) 是由中草藥和薑黃萃取出來的多酚類物質，近年研究指出薑黃素具有抗氧化、抗發炎、抗癌以及預防癌症產生 (chemoprevention) 的能力，然而，薑黃素是如何抑制攝護腺癌細胞侵襲的分子機制尚未清楚了解。在本篇研究裡，我檢測了薑黃素對於攝護腺癌細胞 (PC-3) 的生長、移動與侵襲的影響。實驗結果指出薑黃素可有效地抑制攝護腺癌細胞的生長、移動與侵襲的能力。為了更進一步研究薑黃素抑制攝護腺癌細胞移動與侵襲的分子機制，我檢測了薑黃素對於間質蛋白酶 (Matriptase) 的影響。間質蛋白酶是第二型嵌膜絲胺酸蛋白酶，近年來研究指出不正常活化的間質蛋白酶可能參與了許多癌症的演進，包括攝護腺癌。而實驗結果指出，薑黃素可以降低間質蛋白酶的表現量及活化，但不影響其基因的表現。此外，薑黃素可以促進已活化的間質蛋白酶釋出到細胞外。更進一步地，我們建立了攝護腺癌細胞侵襲能力進化的細胞模式 (PC-3 以及M2I1 PC-3 cells)，並檢驗了薑黃素對此 PC-3 侵襲細胞模式的侵襲能力之影響。 M2I1 細胞比起 PC-3 細胞具有大約多達一倍的侵襲能力。而薑黃素可抑制M2I1細胞侵襲能力，其原因可能是藉由抑制了活化的間質蛋白酶，而非透過影響上皮-間質轉化 (Epithelial-mesenchymal transitions) 的過程。此外，我也檢驗了薑黃素對於過量表達間質蛋白酶的細胞其侵襲能力的影響，透過建立大量穩定表現間質蛋白酶的 CWR22Rv1 細胞，發現大量表現間質蛋白酶的CWR22Rv1細胞可提升高達一倍的侵襲能力。而薑黃素亦可抑制這些大量表現間質蛋白酶的 CWR22Rv1 細胞之侵襲能力，且抑制的現象隨著薑黃素的濃度上升而增加。此外，實驗結果也顯示出薑黃素可以抑制因表皮生長因子 (EGF和Heregulin) 所刺激而增加的PC-3 細胞侵襲能力。整體來說，實驗結果指出薑黃素具有抑制間質蛋白酶功能而達到壓抑攝護腺癌細胞侵襲的能力。

關鍵字

薑黃素 ； 攝護腺癌 ； 侵襲 ； 間質蛋白酶

並列摘要

Prostate cancer is the second leading cause of cancer-related death in men of the western world. In Taiwan, the incidence and mortality of prostate cancer have been rising progressively in recent years. The high mortality of prostate cancer is mainly due to the cancer progression to an androgen-independent state and/or high cancer cell migration, invasion and metastasis. Curcumin, a polyphenol substance derived from the herbal remedy and dietary spice turmeric, has been shown with potent antioxidant, anti-inflammatory, anti-carcinogenic, and chemopreventive effects. However, the molecular mechanism how curcumin suppresses prostate cancer cell invasion is not well understood. In this study, I examined the effect of natural compound curcumin on cell proliferation, migration and invasion in human prostate cancer PC3 cells. The results showed that curcumin could significantly suppress prostate cancer cell proliferation, migration and invasion. To further investigate the molecular mechanism in which curcumin was able to inhibit prostate cancer cell migration and invasion, I examined the effect of curcumin on a tumor-promoting, membrane-anchored serine protease, matriptase, since recent studies have shown that dysregulation of matriptase activation can promote the progression of many cancers including prostate cancer. The data showed that curcumin was able to reduce the activated levels with no effect on its gene expression of matriptase. Moreover, the reduction of activated matriptase by curcumin was partly due to the curcumin promoting the shedding of activated matriptase into extracellular environment. Furthermore, we established a PC-3 cell invasion progression model (parental and M2I1 PC-3 cells) with an increasing cell invasion capability and examined the effect of curcumin on different PC-3 cell invasion. The invasive ability of M2I1 cells was enhanced by approximate one fold, compared to the parental PC-3 cells. Curcumin-decreased M2I1 cell invasion was partly due to suppressing the level of activated matriptase but not via altering EMT process. I also examined the effect of curcumin on the cell invasion of those cells with matriptase overexpresion. I established the stable pools of CWR22Rv1 cells with matriptase overexpression and found that expression of matriptase in CWR22Rv1 cells enhanced their invasive ability approximately one fold. With curcumin treatment, the invasive ability induced by matriptase overexpression was suppressed in a dose-dependent manner. In addition, our data also showed that curcumin was able to suppress EGF- or heregulin-stimulated PC-3 cell invasion. In summary, the data indicated that curcumin exhibits a suppressive effect on prostate cancer cell invasion, at least in part by down-regulating matriptase function.

並列關鍵字

curcumin ； prostate cancer ； invasion ； matriptase

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