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題 名 | 天花粉類抗癌蛋白之研究=Studies on Trichosanthin-Like Antitumor Proteins |
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作 者 | 林榮耀; | 書刊名 | 中醫藥年報 |
卷 期 | 16:2 1998.05[民87.05] |
頁 次 | 頁135-167 |
分類號 | 414.33 |
關鍵詞 | 核醣體; 核醣體抑制蛋白; 免疫毒素; Immunotoxin; Ribosome-inactivating proteins; RIPs; Ribosome; |
語 文 | 中文(Chinese) |
中文摘要 | 天花粉是以前中國鄉村常用的墮胎藥,它是由中國藥用植物 Trichosanthes kirilowii的根部萃取得到。由天花粉分離純化出來的一 種蛋白質被命名為Trichosanthin(TCS),其分子量大約26 kDa,經由實 驗證實具有抗腫瘤(anti-tumor)及抗愛滋病毒(anti-human immunodeficiency virus)的活性。一些報告指,TCS出對於絨毛膜癌 (choriocaracinoma)及黑色素瘤(melanoma)有較強之抑制活性。 Trichosanthes kirilowii屬於葫蘆科植物,在試管內(In Vitro)實驗證 實TCS可以抑制兔子reticulocyte cell free system之蛋白質生合成, 並且具有N-glycosidase 的活性。因此本實驗希望由一些屬於葫蘆科的植 物,如蛇瓜(Trichosanthes anguina)、準人瓜(Sechium edule)等植物中 篩選與TCS類似具有抗癌活性的蛋白質,同時進一步探討其作用機轉。 由蛇瓜、準人瓜種子中純化出具有抑制蛋白生合成活性的蛋白,分別命 名為Trichosanthin-S及Sechiumin,由SDS/PAGE分析二者分子量分別 為35 kDA及28 kDa 且均由單一多胜太所組成。抑制cell-free system 的IC�t分別為0.2 nM,0.8nM;以老鼠肝臟抽出之核醣體為材料測定其 N-glycosidase activity 得到其至0.1 nM均尚可有活性。另外若用完整 的癌細胞分別處理這兩種蛋白,則需5000∼6000nM之濃度才能達到抑制 蛋白質合成的IC�t。對於癌細胞生長或DNA、RNA生合成之抑制則不顯著, 所以由這些結果顯示,Trichosanthin-S及Sechiumin均如同TCS一樣屬 於第一型(typeⅠ)核醣體抑制蛋白(ribosome-inactivating proteins; RIPs)。由於是第一型核醣體抑制蛋白,無法進入完整細胞中所以毒殺完 整細胞能力較差,但對於cell-free system則有很強的抑制蛋白合成效 果,由老鼠肝臟核醣體的N-glycosidase活性測定中,可知這兩種蛋白有 很強的活性,因此接下來可以化學修飾法與其它特異性抗體連接 (cross-link)發展免疫毒素(immunotoxin)以增加對特定癌細胞的毒殺作 用。 |
英文摘要 | Trichosanthin (TCS) was isolated from the Chinese medicinal herb, Tian-hua-fen obtained from root tubers of the Chinese plant, Trichosanthes kirilowii. It possesses anti-tumor and anti-human immunodeficiency virus properties. In vitro, TCS is more active toward choriocarcinomas and melanoma. Trichosanthes kirilowii belongs to the Curcurbitacae family, and so dose Momordica charantia, from which a component similar to TCS was purified. The present investigation is to screen the similar anti-tumor activity as TCS from the members of Curcurbitacae family, for examples, Trichosanthes anguina (蛇 瓜)、Sechium edule Swartz(準人瓜)and to character these biolobical activities. The aims of the study are to obtain these components or their derivaties for medical use such as the treatment of cancers. We purified two new toxic proteins, which we propose to call Trichosanthin-S and Sechiumin, from the seeds of Trichosanthes anguina and Sechium edule Swartz, Cureurbitaceas growing in Taiwan. These two toxic proteins have all the properties of ribosome-inactivating proteins (RIPs) like the A-chains of abrin and the related toxins (ie Trichosanthin). These RIPs catalytically inactivate 60s ribosomal subunits by removing adenine from the position 4324 of 28s rRNA and there by inhibite the protein biosynthesis. From SDS/PAGE, it showes that the molecular weights of Trichosanthin-S and Sechiumin were 35 kDa and 28 kDa respectively. The two toxic proteins inhibite the protein synthesis of a rabbit reticulocyte lysate with an IC�t of 0.2 nM and 0.8nM respectively and have much less effect on protein synthesis of HeLa cells with IC�tof 5000∼6000 nM. After rat liver ribosomes were treated with 0.1 nM Trichosanthin-S or Sechiumin, the total rRNAs were isolated and treated with aniline, and a new rRNA fragment could be clearly observed from PAGE. Trichosanthin-S and Sechiumin belong to type Ⅰ RIPs, which do not possess a lectin domain, do not bind easily to cells, and consequently have a relatively low native cytotoxic activity. with the aim of obtaining selectively toxic molecules, many RIPs have been conjugated to carrier molecules capable of delivering them to specific target cells. Antibodies are the obvious choice for preparing conjugates immunotoxins. Several type I RIPs have been used to prepare immunotoxin (ie gelonin, PAP, saporin...), so we suggeste that Trichosanthin-S and Sechiumin could cross link with specific antibodies to produce highly effective immunotoxins. |
本系統中英文摘要資訊取自各篇刊載內容。