2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38548According 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 ability. Moreover, metastasis may complicate the clinical management and lead to a poor prognosis for cancer patients. The cerebrovascular and heart diseases are the second and third among the top ten of death in Taiwan. Atherogenesis, is considered as the primary clinical outcome of these disease, is characterized by lipid deposition, a chronic inflammatory response, and chronic wound healing processes. Apoptosis may play a role in endothelial cell lining defects, necrotic core formation, or plaque erosion and rupture. Among the variety of proapoptotic factors present in atherosclerotic plaques, oxidized low-density lipoproteins (oxLDLs) are thought to play a crucial role by concomitantly inducing lipid storage, local inflammation, and toxic events. Berberine, an isoquinoline alkaloid, is extensively studied for many years, and has been recently marketed as a dietary supplement in the United States and Europe. It has been approved by the Food and Drug Administration (FDA) and Center for Food Safety and Applied Nutrition (CFSAN), and most research has been conducted on berberine, which has anti-bacteria, anti-biotic, anti-inflammatory, antioxidant and immune-system stimulating properties. Berberine also has the hyperlipidemia and hyperglycemia lowering effects. However, studies on the inhibitory effects of berberine on cancer cell invasion and lung cancer proliferation have been relatively less and warrant a further study. In our preliminary study, cell viability and invasion in the various cancer cells (H1299, HeLa, HS578T, and SK-Hep1) was significantly inhibited by berberine. Berberine also inhibited the secretion of MMP-2 and u-PA by H1299 and HS578T cells, and was sufficient to inhibit tumor growth of H1299 xenografted nude mice. Berberine could against oxLDL-induced cytotoxicity in HUVECs and macrophage. Therefore, in the first year of this study, we propose to (1) investigate the effect of berberine on invasion of various cancer cells and analyze the protein and mRNA levels of invasion relevance genes, (2) investigate the effect of HUVECs by berberine in anti-angiogenesis, (3) analyze the effects for the down-regulation of cell viability and induced apoptosis, (4) investigate the protective effects of berberine on against oxLDL- induced cytotoxicity in endothelial cells and macrophage. In the second year of this study, we aim to explore the underlying mechanism involved in (1) anti- invasiveness, (2) anti-angiogenesis, (3) anti-proliferation, induced apoptosis, and (4) reduced the development of atherosclerosis by berberine. In the third year of this study, we aim to (1) via an LLC-bearing mice model and BALB/c nude mice model determine the effect of berberine on metastasis, (2) analyze the effect of berberine on angiogenesis process in vivo, (3) investigate tumor growth of different human cancer cells in vivo via cancer cell xenografted nude mice model, and (5) finally analyze the effect of berberine on antiatherosclerotic effects in rabbits with experimental atherosclerosis.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且具有異常的增生能力。其中癌細胞 的侵入及轉移亦是癌症導致死亡以及治療複雜度提昇的主要原因。此外，腦血管與心臟 疾病在近幾年來則是分居台灣地區十大死因之第二及第三名，僅次於惡性腫瘤之後。動 脈硬化(atherogenesis)的特徵為脂質沉積、慢性發炎及慢性傷口癒合的過程，最後造成血 管栓塞。細胞凋亡在內皮細胞損傷、壞死的核心形成及血管斑塊斷裂扮演重要角色。動 脈粥狀硬化形成初期主要是經由氧化型低密度脂蛋白在內皮細胞的累積並活化內皮細 胞及增加與單核球(monocytes)間之黏著作用。 黃連素( berberine; 又稱小蘗鹼) 是一種isoquinoline ( 異? ? ) 衍生物 benzylisoquinoline proberberine 型的生物鹼。Berberine 在歐美地區已經被廣泛的應用 在健康食品添加物裡面，且通過了美國藥物及食品管理局(FDA) 及center for food safety and applied nutrition (CFSAN)的認證，其療效包括減緩腹瀉及發炎反應，調控 免疫系統作用、對抗細菌、黴菌，另外berberine 還具有抗氧化、降血糖及減低膽固 醇中的低密度膽固醇。更有研究指出黃連素具有抑制癌瘤細胞增生及生長作用，使 細胞週期停滯並造成細胞的凋謝死亡。然而berberine 對於抑制癌瘤細胞轉移能力的 相關研究則尚未清楚，及其是否會抑制肺癌細胞的生長亦不明瞭。本實驗室初步分 析了berberine 對肺癌(H1299)、乳癌(HS578T)、子宮頸癌(HeLa)與肝癌(SK-Hep1)細 胞侵襲、轉移、增生抑制能力，發現berberine 不僅對肺癌細胞的存活率有明顯抑制 作用，對各種癌瘤細胞侵襲能力抑制效果也非常顯著，並可有效抑制MMP-2 及u-PA 蛋白分解? 的表現；另外，對血管新生作用也具有抑制效果，而在動物模式下也發現有良好的抗腫瘤效果。並且發現berberine 可降低氧化型低密度脂蛋白對血管內皮 細胞與巨噬細胞的傷害。 綜合上述，berberine 在抑制癌瘤細胞侵襲轉移能力研究與預防心血管疾病方面應該 會有所貢獻，所以本計畫第一年擬進行: (1)利用invasion assay、傷口癒合實驗、adhesion assay，探討berberine 抑制多種不同癌細胞轉移、貼附之能力，並分析berberine 對癌瘤 細胞其migration/invasion 相關基因之蛋白層次與mRNA 層次的影響; (2) 以tube formation assay，探討berberine 抑制血管新生之能力，並分析berberine 對HUVECs 的侵 襲能力、細胞存活與血管新生相關之基因表達；(2) 利用MTT assay、flow cytometry、 colony formation、mitochondrial membrane potential assay、DNA fragmentation assay、 TUNEL、Annexin V-conjugated Alexafluor 488，探討berberine 抑制癌細胞增生並誘導癌 細胞凋亡之能力；(4)利用氧化型LDL (oxLDL)處理HUVECs 與macrophage，探討 berberine 是否會抑制由oxLDL 所造成macrophage 與血管內皮細胞之傷害。計畫第二年 擬進行: 利用Western blot、Luciferase assay、大量表達之載體，深入探討berberine 抑制 (1)癌細胞轉移、(2)血管新生、(3)細胞增生、誘導癌細胞凋亡之能力與(4)降低動脈硬化 與保護血管內皮細胞其中的詳細機轉與訊息傳遞路徑。計畫第三年擬進行: (1)免疫缺陷 的小鼠(BALB/c nu/nu mice)由尾靜脈接種癌細胞，探討berberine 抑制癌細胞轉移之能 力；(2)由in vivo 實驗將matrigel 混合VEGF，再處理或不處理berberine，種到C57BL/6 老鼠皮下，再將matrigel 取出，觀察其抑制血管新生之能力；(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癌瘤細胞，腹腔注射berberine，觀察腫瘤 的生長；(4)以紐西蘭白兔餵食高膽固醇食物，並同時餵食berberine，觀察其能否抑制血 管動脈硬化。黃連素抑制癌細胞轉移誘導癌細胞凋亡抑制血管新生抑制動脈硬化berberineinhibit cancer cells metastasisinduce cancer cells apoptosisanti-angiogenesisanti-atherogenesisProject