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Title: 一步合成AS1411修飾的不規則形態金奈米粒子應用於抑制癌細胞生長
Authors: Chiu, Shih-Jen
Contributors: 國立臺灣海洋大學:生命科學暨生物科技學系
Keywords: AS1411;適合體;核仁素;金奈米粒子;癌細胞
AS1411;Aptamers;Nucleolin;Gold nanoparticles;Cancer cells
Date: 2017
Issue Date: 2018-08-22T06:31:17Z
Abstract: AS1411適合體 (aptamer) 是一個富含鳥糞嘌呤 (guanine, G) 所構成的穩定G-四聯體 (G-quadruplex),因其為對核仁素 (nucleolin, NCL) 具有高專一性結合特性並有顯著抑制癌細胞生長之功效。核仁素是一個多功能核仁蛋白,大部分癌細胞膜會有大量表現的核仁素。因此,藉由AS1411和核仁素的專一性結合可將搭載上AS1411的奈米粒子送入細胞中,期望達到標定和抑制癌細胞生長效果。本實驗製備AS1411修飾的金奈米粒子 (gold nanoparticles, Au NPs),簡稱為AS1411Au NPs,是利用一步合成方式將末端帶有poly (A) 的AS1411修飾於4-羥乙基哌嗪乙磺酸 (4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid, HEPES) 等緩衝溶液所合成的不規則Au NPs。不同緩衝溶液如 (3-(N-morpholino)propanesulfonic acid, MOPS)、(3-(N-morpholinyl)-2-hydroxypropanesulfonic acid, MOPSO) 和 (piperazine-1,4-bis(2-hydroxy-3-propanesulfonic acid, POPSO) 可製備不同不規則形狀Au NPs,使用不同AS1411濃度也可控制不規則Au NPs的大小與構形。在初步實驗結果證實AS1411Au NPs可選擇性與乳癌細胞 (MCF-7) 結合。我們期待當AS1411Au NPs 進入癌細胞後因細胞內高濃度穀胱甘肽 (glutathione, GSH) 與Au NPs的金硫鍵結力可使AS1411於Au NPs表面釋放而達到毒殺細胞的效果。在初步實驗結果已證實GSH可導致將球形奈米Au NPs (13 nm) 上的AS1411釋放量達39%。然目前細胞實驗結果發現與未修飾AS1411相比,AS1411Au NPs並無明顯促進抑制癌細胞生長。
AS1411 is a guanine-rich (G-rich) oligodeoxyribonucleotide aptamer, which can form a stable G-quadruplex structure. Studies show that AS1411 can specifically bind to nucleolin (NCL), a multifunctional nucleolar protein located in cell. NCL is overexpressed on cell membrane in most cancer cells. Hence, AS1411-conjugated nanoparticles can be internalized in to the cells via receptor-mediated endocytosis after they specifically interact with nucleolin on the cell membrane, that enables cancer cell labeling and inhibition of cell growth. In this study, we developed a simple one-step method for the synthesis of irregular shaped gold nanoparticles (Au NPs) modified with poly (adenine) -terminal AS1411. The functional Au NPs are prepared through a simple reaction of HAuCl4 with Good's buffers such as (4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid, HEPES), (3-(N-morpholino) propanesulfonic acid, MOPS), (3-(N-morpholinyl)-2-hydroxypropanesulfonic acid, MOPSO) and (piperazine-1,4-bis(2-hydroxy-3-propanesulfonic acid, POPSO) in presence of AS1411. The AS1411 aptamers self-assemble on Au NPs via strong interaction between poly (adenine) and Au NPs. The concentration of AS1411 plays a vital role in controlling the morphology and size of Au NPs. The preliminary results show that AS1411−Au NPs can selectively target MCF-7 breast cancer cells. We believe that, after the AS1411−Au NPs are internalized in to the cells, the intracellular glutathione will induce the AS1411 release from Au NPs, and therfore, they inhibit cancer cell growth. Our results reveal that glutathione can induce the release of about 39% of AS1411 from spherical Au NPs (13 nm). However, AS1411−Au NPs do not show enhanced inhibition effect on cancer cell growth compared to free AS1411. In our future work we will employ AS1411–Au NPs as a cancer drug carrier to specifically target cancer cells and enhance their inhibition properties.
URI: http://ethesys.lib.ntou.edu.tw/cgi-bin/gs32/gsweb.cgi?o=dstdcdr&s=G0010236033.id
Appears in Collections:[生命科學暨生物科技學系] 博碩士論文

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