2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38758According to the announcement of Department of Health, cancer is the biggest death rate among the top ten causes in Taiwan. So far, the most important thing for finding new cancer drug is urgent. It has been reported that cancer cells are not able to go through the normal apoptosis pathway and has abnormal proliferation and metastasis ability. Moreover, metastasis, the major cause of cancer death and various treatment strategies have targeted on preventing the occurrence of metastasis, is a multi-step process involving change of cytoskeleton, cell adhesion and proteolytic degradation of the extracellular matrix (ECM), essential to achieving cell motility. Chinese medicinal herbs have been used for therapeutic purposes in traditional and folk medicine for a long time. A recent report showed that many kinds of Chinese medicinal herbs have the antioxidant, anti-inflammation, anti-allergic and anti-tumor activeness. However, the effects and detailed mechanisms of Chinese medicinal herbs on cancer cell metastasis and inducing cell apoptosis were still unclear. In this study, anti-metastasis and anti-tumor agents were investigated using 50% alcohol extracts of Phyllanthus urinaria L. (PUE) and Psoralea corylifolia L. (PCE), respectively, on various lung cancer call lines (A549, H1299, and LLC) and breast cancer cell lines (MDA-MB-231, HS578T, and JC). In our preliminary study, we demonstrated that PCE induced cell death on MDA-MB-231 cells. PUE inhibited cell invasion and the secretion of MMP-2, and was sufficient to inhibit IL-1β- and PMA-induced MMP-9 expression in A549 human lung cancer cells. PUE also could increase E-cadherin expression in A549 cells. Therefore, in the first year of this study, we propose to (1) investigate the effect of PUE on invasion of various cancer cells and analyze the protein and mRNA levels of invasion relevance genes, (2) investigate the effect of cytoskeleton conformation by PUE in human lung cancer cells, (3) analyze the effects for the down-regulation of cell viability and induced apoptosis by PCE in human breast cancer. In the second year of this study, we aim to explore the underlying mechanism involved in (1) anti-invasiveness, (2) anti-proliferation, induced-apoptosis, and (3) the influences of the cytophysiological changes by PUE and PCE. In the third year of this study, we aim to (1) purchase and identify the active component of PUE and PCE, (1) via an LLC-bearing mice model and BALB/c nude mice model determine the effect of PUE on cancer metastasis, (2) finally investigate tumor growth of different breast cancer cells in vivo via cancer cell xenografted nude mice model and BALB/c-bearing mice model.根據衛生署公佈之台灣十大死亡原因顯示癌症仍為十大死因之首，居高不下，如何治療 癌症病人，尋找更有效的抗癌藥物，是一個迫切的問題。目前有許多文獻指出，癌細胞與正 常細胞差別在於無法進行一般細胞凋亡、具有異常的增生能力與高度轉移能力。而癌細胞的 轉移往往是造成癌症病人病灶復發致死及最難根治的主要原因，其過程包括細胞骨架的改 變、細胞的貼附、細胞外基質的降解及細胞侵襲能力的改變。長期以來，中草藥被應用於傳 統醫學及民俗療法中的各種治療，已有多種中草藥被確認其具有抗氧化、抗發炎、抗過敏及 抗癌等活性，但是到目前為止，對於中草藥抑制癌細胞轉移、誘導癌細胞凋亡以及其中之詳 細機轉的都還不是很清楚。在本實驗中，我們利用葉下珠與補骨脂之酒精(50%)萃取物觀察 其抑制多種肺癌(A549, H1299, LLC)轉移及乳癌細胞(MDA-MB-231、HS578T 與JC) 生長之 功效。經本實驗室初步分析，發現補骨脂之萃取物能有效誘導人類乳癌細胞(MDA-MB-231) 走向死亡的現象。而葉下珠之萃取物則可明顯抑制人類肺癌細胞(A549)之細胞侵襲及MMP-2 蛋白分解酶的表現，此外葉下珠亦可有效抑制癌症促進因子PMA 及IL1-β所誘導之細胞癌化 能力，並顯著促進細胞骨架蛋白E-cadherin 的表達。綜合上述，在抑制癌瘤細胞侵襲轉移能 力研究與促進癌細胞走向凋亡方面應該會有所貢獻，所以本計畫擬分為三年進行，第一年擬 進行: (1)利用invasion assay、傷口癒合實驗、adhesion assay，Superarray 探討葉下珠抑制多 種不同癌細胞轉移、貼附之能力，並分析葉下珠對肺癌細胞其migration/invasion 相關基因之 蛋白層次與mRNA 層次的影響，(2)與其是否會影響多種細胞骨架蛋白的表達；(3)利用MTT assay、flow cytometry、colony formation、mitochondrial membrane potential assay、DNA fragmentation assay、TUNEL、Annexin V-conjugated Alexafluor 488，探討補骨脂抑制乳癌細 胞增生並誘導癌細胞凋亡之能力。計畫第二年擬進行: 利用Superarray、proteomic assay、 Western blot、Luciferase assay、大量表達之載體，深入探討葉下珠抑制(1)由PMA 與IL1-β所 誘導癌細胞轉移、(2)細胞骨架重整與(3)細胞增生、誘導癌細胞凋亡之能力其中的詳細機轉與 訊息傳遞路徑。計畫第三年擬進行: (1)購買文獻已知葉下珠與補骨脂之單離成分，做有效成 分之鑑定分析; (2)免疫缺陷的小鼠(BALB/c nu/nu mice)由尾靜脈接種肺癌細胞，與 LLC-bearing mice model 探討葉下珠抑制癌細胞轉移之能力；(3)以免疫缺陷的小鼠(BALB/c nu/nu mice)皮下接種各種不同之人類乳癌細胞，與JC-bearing mice model，觀察補骨脂抑制 腫瘤生長的能力。癌症轉移細胞凋亡基質金屬蛋白酶細胞骨架蛋白metastasisapoptosisMMPcytoskeletonProject