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全2?-F/OMe-siRNA的设计合成及其新型混合脂材纳米制剂抗肝癌活性评价 被引量:1
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作者 高宇睛 王玺贤 +5 位作者 潘宇飞 汪全鑫 朱月洁 潘德林 关注 杨振军 《药学学报》 CAS CSCD 北大核心 2023年第6期1634-1640,共7页
本文设计合成了多条全2?-F/OMe修饰的抗肝癌siRNA,并在细胞水平进行活性评价,探究2?-F/OMe修饰的位置差异对siRNA活性的影响。使用K&A DNA/RNA H-8合成仪合成全2?-F/OMe修饰siRNA,利用中性胞苷脂材DNCA混合阳离子脂材CLD包载递送si... 本文设计合成了多条全2?-F/OMe修饰的抗肝癌siRNA,并在细胞水平进行活性评价,探究2?-F/OMe修饰的位置差异对siRNA活性的影响。使用K&A DNA/RNA H-8合成仪合成全2?-F/OMe修饰siRNA,利用中性胞苷脂材DNCA混合阳离子脂材CLD包载递送siRNA入胞。通过RT-qPCR实验检测Huh-7和HepG2细胞的靶基因沉默活性, CCK-8实验检测Huh-7和HepG2细胞增殖活力, RNA结合蛋白免疫沉淀及RT-qPCR实验检测siRNA载入Ago2蛋白的情况,流式细胞术检测药物摄取和细胞凋亡, Western blot实验检测PLK1蛋白的表达。部分全2?-F/OMe修饰siRNA增强了与Ago2蛋白的结合,进而增强了基因沉默活性及肝癌细胞增殖抑制活性。此外,多数siRNA修饰物制剂的Huh-7细胞摄取率提高,更大幅度下调PLK1蛋白,诱导更多Huh-7细胞凋亡,其中siPLK1A3最优。以上结果为进一步研究siRNA修饰模式及研发抗肝癌新型制剂提供了工作基础。 展开更多
关键词 小干扰RNA 全2?-修饰 抗肝癌 Ago2 DNCA/cld
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Evaluation of the intracellular trafficking of siRNAs in A375 cells by confocal laser scanning microscopy
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作者 Yiping Diao Jing Sun +3 位作者 Mengyi Yang Bo Xu Lihe Zhang Zhenjun Yang 《Journal of Chinese Pharmaceutical Sciences》 CAS CSCD 2016年第12期859-868,共10页
Investigation intracellular trafficking of siRNAs following their delivery to cells is of great interest to elucidate dynamics of siRNA in cytoplasm. In this study, we present a novel confocal laser scanning microsco... Investigation intracellular trafficking of siRNAs following their delivery to cells is of great interest to elucidate dynamics of siRNA in cytoplasm. In this study, we present a novel confocal laser scanning microscopy (CLSM) method to evaluate a novel delivery system of 3'-peptide-siRNA therapeutic, which was named 3'-pAs-siRNA/CLD. This method could not only calculate the content of the intracellular 3'-peptide-siRNA, but also quantify its co-localization with cellular substructure. We observed that 3'-pAs-siRNA/CLD, which provided the better antitumor capability, also had a better cell uptake, endosome escape and a longer retention time in A375. This novel strategy was proved to be efficient, quantified and visualized, thus making the dynamics research of siRNA in cytoplasm clear and simplified. 展开更多
关键词 sirna/cld complex Endosomal escape Cytoplasmic distribution Confocal laser scanning microscopy
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