適配體介導(dǎo)脂質(zhì)體靶向遞送siRNA的研究
發(fā)布時(shí)間:2018-04-18 05:34
本文選題:適配體 + siRNA ; 參考:《中國(guó)生物工程雜志》2015年01期
【摘要】:為探討適配體介導(dǎo)的脂質(zhì)體靶向遞送siRNA的可行性,采用前列腺癌細(xì)胞膜表面抗原(PSMA)的適配體A10-3.2與脂質(zhì)體結(jié)合,構(gòu)建適配體-脂質(zhì)體靶向遞送體系(Apt-LP),并利用Apt-LP遞送p EGFP-N1質(zhì)粒和Bcl2 siRNA到前列腺癌細(xì)胞LNCa P(PSMA+)和PC-3(PSMA-),轉(zhuǎn)染48h后,用熒光顯微鏡檢測(cè)綠色熒光蛋白的表達(dá),q PCR檢測(cè)Bcl2 mRNA表達(dá),蛋白印記法檢測(cè)Bcl2蛋白表達(dá),Hoechst 33258核染法分析體外抗腫瘤活性。結(jié)果顯示:與脂質(zhì)體遞送體系相比,Apt-LP顯著提高遞送p EGFP-N1質(zhì)粒和Bcl2 siRNA到靶細(xì)胞LNCa P(PSMA+)的效率;顯著提高Bcl2 siRNA誘導(dǎo)的靶細(xì)胞LNCa P(PSMA+)Bcl2基因沉默效應(yīng),更有效的誘導(dǎo)靶細(xì)胞LNCa P(PSMA+)凋亡。結(jié)果表明:Apt-LP是一種有效的siRNA靶向遞送體系,具有潛在的臨床應(yīng)用價(jià)值。
[Abstract]:In order to investigate the feasibility of aptamer mediated liposome targeting delivery of siRNA, the aptamer A10-3.2 of prostate cancer cell membrane surface antigen (PSMA) was used to bind to liposome.Aptamer liposome targeting delivery system (Apt-LPN) was constructed, and Apt-LP was used to deliver p EGFP-N1 plasmid and Bcl2 siRNA to prostate cancer cell line LNCa P(PSMA) and PC-3PSMA-G. After transfection for 48 h, the expression of green fluorescent protein (GFP) and Bcl2 mRNA were detected by fluorescence microscope.The expression of Bcl2 protein was detected by protein imprinting method. Hoechst 33258 nuclear staining method was used to analyze the anti-tumor activity in vitro.The results showed that compared with liposome delivery system, Apt-LP significantly increased the efficiency of delivering p EGFP-N1 plasmids and Bcl2 siRNA to target LNCa P(PSMA, increased the silencing effect of LNCa P(PSMA Bcl2 gene induced by Bcl2 siRNA, and more effectively induced LNCa P(PSMA apoptosis.The results show that: Apt-LP is an effective siRNA targeting delivery system and has potential clinical application value.
【作者單位】: 蘭州交通大學(xué)化學(xué)與生物工程學(xué)院;甘肅省醫(yī)學(xué)科學(xué)研究院;
【基金】:甘肅省中青年基金(1107RJYA033) 蘭州交通大學(xué)校青年基金(2013012)資助項(xiàng)目
【分類號(hào)】:R943
【共引文獻(xiàn)】
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