本文選題：適配體 + 聚陽離子載體�。� 參考：《中國醫(yī)院藥學(xué)雜志》2017年10期

【摘要】：目的:研究適配體SP94修飾的陽離子基因載體H_3R_5,合成新型肝靶向納米復(fù)合物SP94-H_3R_5/miR195,增加對(duì)肝癌細(xì)胞的靶向性,提高基因的轉(zhuǎn)染效率。方法:經(jīng)半胱氨酸修飾的SP94與H_3R_5末端的半胱氨酸發(fā)生氧化反應(yīng),組裝得到SP94-H_3R_5,利用~1 H-NMR鑒定SP94-H_3R_5載體的結(jié)構(gòu),通過電位粒度儀測(cè)定納米復(fù)合物的電位和粒徑,利用瓊脂糖凝膠電泳考察載體對(duì)miR195的壓縮能力。以體外培養(yǎng)的人肝癌SK-Hep-1為研究對(duì)象,CCK8法檢測(cè)SP94-H_3R_5和H_3R_5對(duì)細(xì)胞增殖的抑制作用,采用激光共聚焦顯微鏡考察肝癌細(xì)胞對(duì)納米復(fù)合物的攝取,以pEGFP為報(bào)告基因考察基因轉(zhuǎn)染效率,Western blot實(shí)驗(yàn)檢測(cè)SK-Hep-1細(xì)胞VEGF的蛋白表達(dá)。結(jié)果:SP94-H_3R_5具有生物相容性,可以壓縮miR195形成穩(wěn)定的納米復(fù)合物,SP94-H_3R_5/miR195與H_3R_5/miR195相比可以更多地被SK-Hep-1攝取(P0.01),SP94-H_3R_5轉(zhuǎn)染效率高于非靶向載體H_3R_5,對(duì)VEGF的阻滯作用也更高(P0.01)。結(jié)論:SP94-H_3R_5兼具納米材料的被動(dòng)靶向作用和適配體的主動(dòng)靶向作用,有潛力成為肝癌治療中的新型載體。
[Abstract]:Aim: to study the cationic gene vector H3RSTE5 modified by aptamer SP94 and synthesize a novel liver targeting nanocomplex SP94-H3R5 / miR195 to increase the targeting of hepatoma cells and improve the efficiency of gene transfection. Methods: the cysteine modified SP94 reacted with cysteine at the end of H_3R_5, and SP94-H3R5 was assembled. The structure of SP94-H_3R_5 carrier was identified by H-NMR. The potential and particle size of the nanocomposites were measured by potentiometric particle size analyzer. Agarose gel electrophoresis was used to investigate the compression ability of the carrier to miR195. The inhibitory effects of SP94-H_3R_5 and H_3R_5 on the proliferation of human hepatocellular carcinoma (SK-Hep-1) in vitro were detected by CCK8 method. The uptake of nano-complexes in hepatoma cells was investigated by confocal laser microscopy. The efficiency of gene transfection was evaluated by using pEGFP as reporter gene. Western blot assay was used to detect the protein expression of VEGF in SK-Hep-1 cells. Results compared with H_3R_5/miR195, the transfection efficiency of S / SP94-H3RS5 was higher than that of H_3R_5/miR195, and the transfection efficiency was higher than that of non-target carrier HNS3R5. The effect on VEGF was also higher than that on VEGF. Conclusion Snx SP94-H3RSP 5 has both passive targeting effect of nano-materials and active targeting of aptamers, and it has the potential to become a new type of carrier in the treatment of liver cancer.
【作者單位】： 上海交通大學(xué)附屬第一人民醫(yī)院臨床藥學(xué)科;第二軍醫(yī)大學(xué)長海醫(yī)院藥學(xué)部;
【基金】：國家自然科學(xué)基金資助項(xiàng)目(編號(hào):81302212) 上海市自然科學(xué)基金資助項(xiàng)目(編號(hào):16ZR1428000) 上海交通大學(xué)醫(yī)工交叉資助項(xiàng)目(編號(hào):YG2014MS32,YG2015QN14)
【分類號(hào)】：R450;R735.7

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