二十二碳六烯酸與二十碳五烯酸對調控SKOV-3人類卵巢腺癌細胞移行及侵襲相關蛋白表現之影響

Effects of docosahexaenoic acid and eicosapentaenoic acid on the expression of migration- and invasion-related proteins in SKOV-3 human ovarian adenocarcinoma cells

林宸宇(Chen-Yu Lin)

指導教授 ： 劉德中

中山醫學大學/健康管理學院/營養學系碩士班/碩士(2015年)

https://doi.org/10.6834/CSMU.2015.00167

摘要

卵巢癌為女性第二常見之癌症，癌細胞轉移是導致卵巢癌患者死亡主要原因。研究證實，二十碳五烯酸(eicosapentaenoic acid, EPA)及二十二碳六烯酸(docosahexaenoic acid, DHA)可降低前列腺癌細胞、肝癌細胞之轉移。本實驗室先前已證實DHA處理MCF-7乳癌細胞後，可抑制12-O-tetradecanoylphorbol-13-acetate (TPA)所誘發之基質金屬蛋白酶-9(matrix metalloproteinases-9, MMP-9)表現及癌細胞移行和侵襲。然而DHA及EPA是否亦可抑制SKOV-3卵巢腺癌細胞之移行及侵襲，其相關作用機制尚不清楚。本研究以SKOV-3卵巢腺癌細胞為實驗模式，探討DHA及EPA對癌細胞轉移作用之影響及其差異。先前研究證實，氧化壓力誘導生長抑制因子-1 (oxidative stress induced growth inhibitor 1, OSGIN-1)和第一型血紅素氧化酶(heme oxygenase-1, HO-1)大量表現與負向調控MMPs表現和活性進而抑制癌細胞生長及轉移有關。本研究預先以各種不種脂肪酸- stearic acid (SA)、oleic acid (OA)、arachidonic acid (AA)、linoleic acid (LA)、γ-linoleic acid (GLA)、α-linolenic acid (LNA)、EPA及DHA處理細胞，結果顯示，僅有DHA可大量誘發OSGIN-1及HO-1蛋白質及mRNA表現。100 μM DHA可抑制SKOV-3卵巢腺癌細胞MMP-9及MMP-2蛋白表現，但只可降低MMP-2酵素活性；而100 μM EPA 可同時抑制MMP-9及MMP-2蛋白表現及其酵素活性。利用傷口癒合試驗(wound-healing assay)及細胞侵襲試驗(invasion)比較DHA和EPA對SKOV-3之移行及侵襲之影響，發現DHA及EPA皆可抑制癌細胞移行及侵襲。此外，利用si-RNA靜默OSGIN-1基因表現後，並無影響由DHA及EPA所抑制MMP-9及MMP-2之表現。綜合上述結果，DHA及EPA皆可抑制SKOV-3卵巢腺癌細胞本身MMP-9及MMP-2之表現，特別是透過降低MMP-2之表現及酵素活性，進而抑制SKOV-3卵巢腺癌細胞之移行及侵襲。此外，DHA抑制MMP-9及MMP-2表現、活性及癌細胞移行並非透過OSGIN-1所調控；而DHA抑制MMP-9及MMP-2之蛋白表現、酵素活性及癌細胞移性可透過HO-1進行調控。

關鍵字

卵巢癌 ； 二十二碳六烯酸 ； 二十碳五烯酸 ； 氧化壓力誘導之生長抑制因子 ； 第一型血紅素氧化酵素 ； 基質金屬蛋白酶癌細胞轉移

並列摘要

Ovarian cancer is the second most common cancer in women worldwide. Metastasis is the major cause of death from ovarian cancer. Previous studies showed that metastasis of various cancer cells, such as prostate cancer and hepatoma cells, were suppressed by eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Our previous study indicated that 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced MCF-7 breast cancer cells migration and invasion were suppressed by DHA though down-regulation of matrix metalloproteinases-9 (MMP-9). However, the inhibitory effects of EPA and DHA on metastasis in ovarian cancer are still not clarified. In the present study, SKOV-3 ovarian adenocarcinoma cells were used to distinguish the anti-metastatic potential between EPA and DHA and the possible mechanisms involved. Induction of oxidative stress induced growth inhibitor 1 (OSGIN-1) and heme oxygenase-1 (HO-1) is negatively related with MMPs-mediated growth and metastasis of cancer cells. In our preliminary data, the protein and mRNA levels of OSGIN1 as well as HO-1 were dramatically induced only in DHA treated group, but other fatty acids including stearic acid (SA), oleic acid (OA), arachidonic acid (AA), linoleic acid (LA), γ-linoleic acid (GLA), α-linolenic acid (LNA) and EPA. We found that 100 μM DHA inhibit MMP-9 and MMP-2 expression and MMP-2 enzyme activity, but both of protein expression and enzyme activity of MMP-9 and MMP-2 were inhibited by EPA in SKOV-3 cells. Furthermore, the cell migration and invasion were suppressed by DHA and EPA. OSGIN-1 si-RNA knockdown could not affect MMP-9 and MMP-2 expression in SKOV-3 cells. These results indicate that suppression of migration and invasion by DHA and EPA is likely through an inhibition of MMP-9 and MMP-2 expression and enzyme activity, especially MMP-2 in SKOV-3 cells. Moreover, the inhibitory effects of DHA on metastasis of SKOV-3 ovarian adenocarcinoma cells via modulated of not OSGIN-1 but HO-1 expression.

並列關鍵字

SKOV-3 ； DHA ； EPA ； OSGIN-1 ； HO-1 ； MMP-9 ； MMP-2 ； Metastasis

參考文獻

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