扁柏酚 對小鼠黑色素瘤B16-F10细胞的抑制作用之機轉探討

黑素瘤是最具侵襲性的癌症之一，對化療具有高度抗藥性，並且其死亡率在全世界正迅速增加當中。腫瘤的入侵和轉移是惡性腫瘤患者治療失敗的主要原因。細胞外基質的分解會通過基質金屬蛋白酶的產生來促進癌細胞的侵入和轉移。根據研究顯示，天然膳食的抗氧化劑似乎能夠抑制腫瘤癌細胞的生長。扁柏酚（Hinokitiol，又名檜木醇），是一種天然的生物活性化合物，最早被發現於台灣扁柏當中，目前早已被用於食品，化妝品和髮油中作為抗微生物劑來使用。根據文獻報導，扁柏酚它還可以抑制血小板聚集和血管平滑肌細胞的增殖。我們目前的研究是，嘗試去評估扁柏酚的抗腫瘤作用，和調查扁柏酚對轉移性黑色素瘤細胞瘤B16-F10細胞的侵襲和遷移有可能的作用機轉，其中基質金屬蛋白酶-1被學者發現在黑色素瘤細胞瘤中會過度表現，而基質金屬蛋白酶-2和9則主要是在黑色素瘤細胞瘤的轉移過程中也會過大量表現，然而在體外實驗中有學者發現，它可能會透過調節腫瘤侵入因子基質金屬蛋白酶和抗氧化酶等來影響黑色素瘤細胞瘤的轉移。利用扁柏酚處理過之後，我們發現會顯示出扁柏酚對小鼠黑素瘤B16-F10細胞的遷移具有濃度依賴性的抑制作用。扁柏酚看起來是可以透過降低基質金屬蛋白酶-1/2/9的活性和表達來獲得以下這些效果：通過阻斷促分裂原活化蛋白激酶（MAPK）信號級聯，包括ERK1/2，p38 MAPK和JNK的磷酸化;通過逆轉降解IκB-α的抑製劑，和抑制參與B16-F10黑素瘤細胞的轉錄因子p65 NF-κB與c-Jun激活的磷酸化;也會通過抑制p65 NF-κB從細胞質向細胞核移位以減少NF-κB的活化。此外，扁柏酚它還會減少芬頓反應所誘導的羥基自由基的產生，並且增強抗氧化酶過氧化氫酶和超氧化物歧化酶在B16-F10黑色素細胞瘤中的活性。這些發現與我們在體外實驗室的結果是相一致性的，另外根據我們在小鼠體內的研究顯示，經過扁柏酚注射治療實驗小鼠過後，的確可以發現扁柏酚會顯著降低小鼠肺轉移結節的總數，回復其肺部的彈性纖維和膠原纖維之分佈，並且會顯著改善B16-F10黑色素細胞瘤中肺泡的組織學改變包括減少肺部的纖維化。考慮到這些研究結果，我們建議可以用扁柏酚來處理黑色素細胞瘤是會顯著阻斷B16-F10細胞的轉移，其作用機轉最有可能是通過抑制基質金屬蛋白酶-1/2/9的激活，抑制MAPK信號級聯，阻止轉錄因子NFk-B和c-Jun的激活，減少羥基自由基形成，以及促進抗氧化酶-超氧化物歧化酶和過氧化氫酶來參與阻斷其癌細胞的遷移。因此，扁柏酚可以是有效的抗癌候選藥物，並且這些結果可用於開發針對惡性腫瘤的新型治療藥劑。

關鍵字

黑素瘤；扁柏酚；基質金屬蛋白酶

並列摘要

Melanoma is one of the aggressive cancers, highly resistive to chemotherapy and its mortality rate is going up speedily worldwide. Invasion and metastasis are the major causes of treatment failure in patients with malignant tumor. The breakdown of extracellular matrix promotes the invasion and migration of cancer cells through the production of matrix metalloproteinases (MMPs). The studied research has revealed that natural dietary antioxidants are able to inhibit the growth of tumor cells. Hinokitiol, a natural bioactive compound found in Chamaecyparis taiwanensis, has been used in food, cosmetics, and hair tonics as an antimicrobial agent. It also has been reported to inhibit platelets aggregation and vascular smooth muscle cell proliferation. Our present study is tried to evaluate the antitumor effects and to investigate the possible mechanisms of action of hinokitiol on invasion and migration against the metastatic melanoma cell line, B16-F10 cells, wherein MMP-1 is dug out to be over expressed and MMPs-2/9 are predominantly convoluted in the metastasis process, by modulating tumor invasion factors MMPs, antioxidant enzymes in vitro. Treatment with hinokitiol demonstrated a concentration dependently suppression of the mouse melanoma B16-F10 cells migration. Hinokitiol looked like to attain this effect by decreasing the activity and expression of MMPs-1/2/9, via blocking the phosphorylation of mitogen activated protein kinase (MAPK) signaling cascades, including extracellular signal-regulated kinase (ERK) 1/2, p38 MAPK and c-Jun N-terminal kinases (JNK); through reversing the degradation of inhibitor of kappa B-α (IκB-α) and suppressing the phosphorylation of transcription factors p65 nuclear factor kappa B (NF-κB) and c-Jun activation involved in B16-F10 melanoma cells; also by inhibiting the translocation of p65 NF-κB from cytosol to nucleus in order to reduce the activation of NF-κB. Furthermore, It also lessened the production of Fenton reaction induced hydroxyl radical (OH•) and enhanced the activities of antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD) in B16-F10 melanoma cells. Compatible with our in vitro laboratory findings, the in vivo study displayed that hinokitiol treatment significantly decreased the total number of mouse lung metastatic nodules, recovered the elastic fibers (EF) and collagens distribution, ameliorated histological alterations and decreased the fibrosis in the lungs of B16-F10 cells injected experimental mice. Considering with these research findings, we suggest that melanoma B16-F10 cells treated with hinokitiol significantly blocked the metastasis, most probably by inhibiting MMPs-1/2/9 activation, suppressing MAPK signaling cascades, preventing transcription factors NF-κB and c-Jun activation, decreasing OH• formation, and promoting antioxidant enzymes SOD and CAT involved in cancer cell migration. Therefore, hinokitiol may be a potent anticancer candidate and these results may be useful to develop the novel therapeutic agents for the malignant tumors.

並列關鍵字

Melanoma ； Hinokitiol ； Matrix metalloproteinases

參考文獻

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扁柏酚對小鼠黑色素瘤B16-F10细胞的抑制作用之機轉探討

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