球薑酮透過NFκB及Hedgehog訊息傳遞抑制乳癌細胞株生長的機制探討

乳癌在台灣婦女癌症中好發率居高不下，先前研究已發現球薑酮可抑制乳癌細胞生長，因此本研究以人類乳癌細胞株MDA-MB-435S與MDA-MB-468為實驗模式探討球薑酮影響乳癌細胞生長的路徑。本實驗給予乳癌細胞不同濃度之球薑酮（MDA-MB-435S為0、10、20、50 μM及50 μM加上40 nM PMA (Phorbol 12-myristate 13-acetate)；MDA-MB-468為0、10、20、40 μM及40 μM加上40 nM PMA）。根據先前文獻指出，細胞核轉錄因子NFκB及Hedgehog（Hh）訊息相關蛋白皆與細胞生長相關，缺氧誘導因子HIF-1α則是參與細胞增生與轉移作用進而影響癌症的進展。本實驗利用西方墨點法分析NFκB、HIF-1α及Hh訊息傳遞路徑相關蛋白之表現量。結果顯示，球薑酮在長時間48小時下給予最高劑量能抑制兩株乳癌細胞NFκB的活性，本篇研究亦偵測p-NFκB的表現量，由結果可看出p-NFκB表現量在兩株細胞亦是受到了抑制。另一表現量HIF-1α在本實驗並未受到球薑酮所影響。進一步探討與Hh訊息傳遞路徑相關之蛋白表現量，發現在48小時之球薑酮最高濃度可抑制Sonic Hedgehog（Shh）、Gli-1、Gli-2活化型與Gli-3活化型的表現；促進Smoothened（Smo）的表現，但對於Patched（Ptch）之表現量並無影響。本篇結果顯示，球薑酮可透過NFκB及Hedgehog訊息傳遞路徑而抑制乳癌細胞株生長。

In Taiwan, the incidence of breast cancer continues to rise. Inhibitory effect of breast cancer cell growth was found by the treatment of Zerumbone in the previous studies. For further investigation of how Zerumbone influence the subsequent signaling pathways and mechanisms after the treatment, we adopted two breast cancer cell lines MDA-MB-435S and MDA-MB-468 as the experiment model. In this study, cell lines were treated with various concentrations of Zerumbone (0, 10, 20, 50 and 50 μM plus 40 nM PMA as NFκB-positive controls to to μM MDA-MB-435S; 0, 10, 20 μM, 40 μM and 40 μM plus 40 nM PMA to MDA-MB-468). In our preliminary studies, transcription factor NFκB are related to Hedgehog protein expression regarding cancer cell growth, and HIF-1α involved in human cancer progression, in terms of cell proliferation and metastasis. Therefore we analyze NFκB, HIF-1α and Hedgehog signaling pathway by western blot technique. The results showed that, NFκB expression was reduced after 48 hours treatment of Zerumbone in the groups receiving high concentration of Zerumbone treatment. While HIF-1α expression pattern remained unchanged, reduced expression of phosphorylation of NFκB was found in both cell lines. Contradicting effect of Zerumbone in Hedgehog signaling pathway was observed, meaning the expression Sonic Hedgehog (Shh), Gli-1, and the active forms of Gli-2 and Gli-3 in the high concentration was inhibited after 48 hours treatment, but it enhanced the expression of Smoothened (Smo). However, Patched (Ptch) expression remained unchanged by Zerumbone. Our results indicate that, Zerumbone is able to inhibit breast cancer cell growth though NFκB and Hedgehog signaling pathways.