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系統識別號 U0026-0812200910381217
論文名稱(中文) 人蔘皂Rh2抗癌活性的研究
論文名稱(英文) An Investigation on the Anticancer Activities of Ginsenoside Rh2
校院名稱 成功大學
系所名稱(中) 藥理學研究所
系所名稱(英) Department of Pharmacology
學年度 91
學期 2
出版年 92
研究生(中文) 陳悅穎
研究生(英文) Yueh-Ying Chen
學號 s2690109
學位類別 碩士
語文別 中文
論文頁數 74頁
口試委員 口試委員-簡伯武
指導教授-李益謙
口試委員-鄭瑞棠
中文關鍵字 癌症  人蔘皂Rh2  血管新生  轉移  肝癌 
英文關鍵字 angiogenesis  metastasis  hepatoma  cancer  Ginsenoside Rh2 
學科別分類
中文摘要 本研究為探討一般公認具有滋養補氣血之人蔘(Panax ginseng CA Meyer)其中主要的抗癌成分—人蔘皂Rh2(Ginsenoside Rh2, G-Rh2) —是藉由何種抗癌分子機制來達成預防與治療癌症的效果。首先,分別測試G-Rh2以及含有1% G-Rh2之中草藥複方—Sabonin—是否對癌細胞的生長具有抑制效果。我們發現Sabonin對於NIH 3T3 /ras 7-4及SW480細胞的生長,具有劑量遞增性的抑制效果。而G-Rh2對於A549、NIH 3T3 /ras 7-4、B16F10細胞的生長,亦具有劑量遞增性的抑制效果。但在相同劑量下,G-Rh2對NIH 3T3及PC-3細胞的生長較不具抑制效果,顯示G-Rh2對癌細胞的生長抑制效果,是具有選擇性的。由流式細胞分析儀測試的結果得知,G-Rh2可使A549、B16F10細胞之細胞週期先停頓在per G0/G1 phase,再進一步促進細胞發生凋亡。我們接著測試G-Rh2對血管新生是否具有抑制效果。已知腫瘤的轉移有賴於血管新生。腫瘤周圍新血管的生長,是提供癌細胞進入血液循環及轉移到遠端位置的良好管道。在細胞移行分析(cell migration assay)實驗中,發現G-Rh2對BAEC及PC-3細胞的移行能力具有劑量遞增性的抑制效果。在雞胚胎絨毛膜血管形成試驗(chorioallantoic membrane assay)中,無法觀察出G-Rh2對血管新生的影響,但 Sabonin萃取液卻可以抑制雞胚胎血管新生。Sabonin亦可以抑制由 NIH 3T3 /ras 7-4細胞誘發的Balb/c小鼠皮下腫瘤。然而,利用C57/BL6小鼠尾靜脈注射黑色素癌細胞B16F10細胞的肺臟轉移模式,發現Sabonin並不具有抑制癌細胞轉移的能力。若以預防的角度來探討,長期於飼料中混合0.1% Sabonin粉末投予B6C3F1小鼠,確實可以預防由致癌劑2-acetylaminofluorence誘導的自發性原位肝癌產生,故再次證實了Sabonin可以抑制體內腫瘤的成長。未來仍需更進一步釐清G-Rh2在癌症發展的各個過程中參與的調控機制,及Sabonin中個別成份在抗癌上扮演的角色,以暸解G-Rh2及Sabonin的個別成份,在腫瘤抑制上更詳細的分子藥理學作用。
英文摘要 The anticancer activity of a pure ginsenoside from Panax ginseng C.A. Meyer, G-Rh2, was investigated. We first tested the cytotoxicities of G-Rh2 and a mixture format termed “Sabonin” that contains 1% G-Rh2 and other herbal ingredients on a variety of cancer cell lines. Sabonin exhibits a dose-dependent growth inhibitions on NIH3T3/ras and SW 480 cells. G-Rh2 exhibits a dose-dependent growth inhibitions of A549, NIH3T3/ras and B16F10 cells. However, the same dose has little effect on NIH3T3 wild-type and PC-3 cells, suggesting discriminatory inhibitions of ginsenoside. Cell flowcytometry assay shows that G-Rh2 arrests cells at the per-G0/G1 phase, and later leads to apoptosis of B16F10 and A549 cells. We then tested the antiangiogenesis effect of G-Rh2. Tumor metastasis is angiogenesis dependent. The new blood vessels embedded in a tumor provide a gateway for tumor cells to enter the circulation and to metastasize to distants sites. Cell migration assay using Boyden chamber shows that G-Rh2 inhibits the movements of bovine aortic endothelial and PC-3 cells. However, in the semi in vivo chicken chorioallantonic membrane (CAM) assay, Sabonin does but G-Rh2 does not inhibit vascular formation. Sabonin when fed orally to Balb/c mouse inhibits strongly the growth of tumor induced by the subcutaneous transplantation of NIH3T3/ras cells into the testing animal. However, Sabonin does not reduce the numbers of tumor colonies induced by the tail vein injection of B16F10 cells into C57BL/6 mouse in the experimental metastasis assay. Feeding food containing 0.1% Sabonin to B6C3F1 mouse inhibits strongly the growth of hepatoma induced by the 2-acetylaminofluorence injected intraperitoneally, suggesting again the effectiveness of Sabonin in inhibiting tumor growth in vivo.
論文目次 目錄....................................................I
圖目錄................................................III
表目錄.................................................IV
中文摘要................................................V
英文摘要...............................................VI
縮寫表................................................VII
第一章研究背景.........................................1
第二章研究目的........................................14
第三章實驗材料........................................16
第一節細胞株..........................................16
第二節試劑............................................16
第四章實驗方法........................................18
第一節離體試驗........................................18
第一段細胞培養........................................18
第二段離體細胞毒性....................................20
第三段合併藥物細胞毒性................................23
第四段細胞凋亡之分析..................................24
第五段癌細胞移行試驗..................................25
第六段內皮細胞移行試驗................................27
第二節活體試驗........................................28
第一段雞胚胎絨毛膜血管形成試驗........................28
第二段癌細胞誘發皮下原位腫瘤模式......................30
第三段癌細胞誘發肺臟轉移腫瘤模式......................31
第四段致癌劑誘發肝臟原位腫瘤模式......................32
第五章實驗結果........................................34
第六章討論............................................42
參考文獻..............................................49
圖表..................................................55
作者簡歷..............................................74
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