- 學位論文
探討白花前胡素抑制人類肝癌細胞侵襲與轉移能力
Investigation of Praeruptorin Inhibits the Migration and Invasion in Human Hepatocellular Carcinoma Cells
摘要
白花前胡素存在於 Peucedanum praeruptorum Dunn 的干燥根中，具有白花前胡甲素 ( Praeruptorin A; PA )、白花前胡乙素 ( Praeruptorin B; PB )、白花前胡丙素 ( Praeruptorin C; PC ) 三種活性成分。之前實驗證明praeruptorum 的活性成分具有神經保護、抗炎和抗癌作用。因此本研究利用白花前胡甲素三種活性成分 ( PA、PB、PC ) 對人類肝癌細胞的生長和轉移的功能及分子機轉。我們證實利用四株人類肝癌細胞(SK-Hep-1、Huh-7、HepG2和 PLC/PRF/5/PRF/5)以MTT、群落形成實驗和細胞週期測定證實PA、PB和PC不會抑制肝癌細胞生長、細胞週期改變且無細胞毒性。此外，我們發現PA會抑制SK-Hep-1和Huh-7細胞轉移和侵襲作用，但是PB和PC則無影響。之前文獻得知ADAM9在肝癌轉移過程扮演重要角色，且TCGA資料庫也證實ADAM9在肝癌組織表現量高於正常組織，ADAM9與腫瘤期數和病人存活率有正相關。我們利用西方墨點法證實ADAM9在SK-Hep-1和Huh-7細胞表現量高於HepG2和 PLC/PRF/5/PRF/5細胞，接著PA處理SK-Hep-1細胞發現明顯抑制ADAM9蛋白表現。我們也證實PA增加p-MEK/p-ERK蛋白表現增加。此外，利用TPA處理HepG2細胞增加細胞轉移和侵襲作用，PA共同處理TPA會明顯抑制HepG2細胞轉移和侵襲能力。綜合以上實驗結果說明 PA抑制肝癌細胞轉移作用可能是透過ADAM9蛋白，未來會提供更多實驗證據說明 PA在肝癌轉移的角色及作用機制。
並列摘要
Praeruptorin was discovered in the dry roots of Peucudanum praeruptorum Dunn, with three types of active ingredients praeruptorin A (PA), praeruptorin B (PB) and praeruptroin C (PC). Previous studies have proved that the active ingredients of praeruptorum possessed neuroprotective, anti-inflammation and anti-cancer activities. In this regard, we implemented three different types of active ingredients of praeruptorin (PA, PB and PC) to study its effects on the proliferation and metastasis of human hepatocellular carcinoma (HCC) cells and its underlying molecular mechanism. With four different types of HCC cells (SK-Hep-1, Huh-7, HepG2 and PLC/PRF/5), our MTT assays, colony formation assays and cell cycle analysis showed that PA, PB and PC did not inhibit the growth of HCC cells, nor altered the cell cycles and had no cytotoxicity against HCC cells. However, we found out that PA can inhibit the migration and invasion ability of SK-Hep-1 and Huh-7 cells, while PB and PC have no effects. ADAM9 have been known to play an important role in the metastasis of HCC. Furthermore, TGCA database analysis also showed that the expression of ADAM9 is higher in HCC tissue than in normal tissue, and have a positive correlation with the tumor stage and the survival of patients. Our western blot analysis showed that SK-Hep-1 and Huh-7 have higher ADAM9 expression than HepG2 and PLC/PRF/5, and PA treatment significantly inhibited the expression of ADAM9 in SK-Hep-1 cells. We also showed that PA treatment increased the expression of p-MEK/p-ERK. In addition, when using TPA to enhance the migration and invasion of HepG2 cells, the addition of PA with TPA will significantly inhibit the migration and invasion of HepG2 cells. Taken together, our data showed that PA may inhibit the metastasis of HCC cells through ADAM9. In the future, we will provide further experimentation results to elucidate the role and underlying mechanism of PA in HCC metastasis..