Functionalizing Liposomes with anti-CD44 Aptamer for Selective Targeting of Cancer Cells
Functionalizing Liposomes with anti-CD44 Aptamer for Selective Targeting of Cancer Cells
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DOI：
10.1021/bc5004313
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发表时间：
2015-07-01
影响因子：
4.7
通讯作者：
Fattal, Elias
中科院分区：
文献类型：
--
作者：
Alshaer, Walhan;Hillaireau, Herve;Fattal, Elias
发现CD 44受体蛋白在许多肿瘤中过表达，并被鉴定为常见的肿瘤干细胞表面标志物之一，包括影响结肠、胰腺、头颈部的肿瘤，使其成为治疗靶向的有吸引力的受体。在本研究中，使用巯基-马来酰亚胺点击反应将先前针对CD 44选择的含2 '-F-嘧啶的RNA适体（Apt 1）成功地缀合至PEG化脂质体的表面。通过位点和zeta电位的变化以及通过琼脂糖凝胶电泳上的迁移来证实Apt 1与脂质体表面的缀合。与游离Aptt相比，Apt 1的结合亲和力在缀合后得到改善。使用两种CD 44（+）细胞系，人肺癌细胞（A549）和人乳腺癌细胞（MDA-M.13-231），以及CD 44（-）细胞系，小鼠胚胎成纤维细胞（NIH/3 T3），通过流式细胞术和共聚焦成像测试Aptl-Lip的细胞摄取。结果表明，与空白脂质体（Mal-Lip）相比，Aptl-Lip的灵敏度和选择性更高。最后，我们证明了抗-C44适体与脂质体表面的成功结合以及Aptl-Lip与表达CD 44的癌细胞的结合偏好，并得出结论，Aptl-Lip作为特异性药物递送系统的有希望的效力。
CD44 receptor protein is found to be overexpressed by many tumors and is identified as one of the Moist common cancer stem, cell surface markers including tumors affecting colon, brat, pancreas, and head and neck, making this an attractive receptor for therapeutic targeting. In this study, 2'-F-pyrimidine-containing RNA aptamer (Apt1); previously selected against CD44, was successfully conjugated to the surface of PEGylated liposomes using the thiol-maleimide click reaction. The conjugation of Apt1, to the surface of liposomes was confirnied by the change in site and zeta potential and by migration on agarose gel electrophoresis. The binding affinity of Apt1 was improved after Conjugation compared to free-Aptt. The cellular uptake for Aptl-Lip was tested by flow cytometry and confocal imaging using the two CD44(+) cell lines, human lung cancer cells (A549) and human breast cancer cells (MDA-M.13-231), and the CD44(-) cell line, mouse embryonic fibroblast cells,(NIH/3T3). The results showed higher sensitivity and selectivity for Aptl-Lip compared to the blank liposomes (Mal-Lip). In conclusion; we demonstrate a successful conjugation of anti-C44 aptarner to the surface of liposome and binding preference of Aptl-Lip to CD44-expressing cancer cells and conclude to,a promising potency of Aptl-Lip as a specific drug delivery system.