整合素配体RGD修饰的阳离子脂质体作为siRNA输送载体的体外研究

整合素配体RGD修饰的阳离子脂质体作为siRNA输送载体的体外研究

杨世进, 杨丽娟, 姜娟, 王坚成^*, 张强

北京大学药学院药剂学系, 北京 100083

收稿日期:2008-03-06 修回日期:2008-05-10 出版日期:2008-06-15 发布日期:2008-06-15
通讯作者: 王坚成*

In vitro investigation of RGD-modified stabilized cationic liposomes as non-viral vehicle for siRNA delivery

Shi-Jin Yang, Li-Juan Yang, Juan Jiang, Jian-Cheng Wang^*, Qiang Zhang

Department of Pharmaceutics, School of Pharmaceutical Sciences, Peking University, Beijing 100083, China

Received:2008-03-06 Revised:2008-05-10 Online:2008-06-15 Published:2008-06-15
Contact: Jian-Cheng Wang*

摘要/Abstract

摘要：

本研究构建了能够靶向肿瘤新生血管的RGD修饰阳离子脂质体(RGD-Lipo), 作为靶向耐药相关基因MDR1的siRNA输送载体并评价其相关的药剂学性质。该脂质体与siRNA形成的复合物粒径控制在200 nm以内,并且对其中所包载的siRNA具有一定的保护作用。体外实验结果表明, 经RGD修饰的脂质体(RGD-Lipo)细胞粘附能力显著增强, 并可增加细胞内siRNA的转染效果。与利用前插法进行靶向修饰相比, 利用后插法进行RGD修饰可有效地改善siRNA的溶酶体释放效率。细胞毒试验结果表明, 后插法制备的pRGD-Lipo-siRNA能够有效逆转人卵巢癌SKVO3/A细胞的耐药性, 并增加阿霉素药物在细胞内的蓄积。体外研究结果证实, 使用pRGD-Lipo-siRNA有利于提高耐药细胞对化疗药阿霉素的敏感度, 并将有可能应用于临床耐药肿瘤的治疗。

关键词: siRNA, siRNA, siRNA, 阳离子脂质体, 阳离子脂质体, 阳离子脂质体, DC-Chol, DC-Chol, DC-Chol, 整合素RGD, 整合素RGD, 整合素RGD, , ,

Abstract:

In this study, the novel RGD-modified stabilized cationic liposomes were developed as the delivery vehicle for siRNA targeting human MDR1 gene. The complex of cationic liposomes and siRNA, RGD-Lipo-siRNA, was prepared with a narrow size distribution below 200 nm. It was shown that the encapsulated siRNA in the liposomes could be effectively protected from serum degradation. Also, enhanced cell binding and intracellular uptake of siRNA in the doxorubicin-resistant human ovarian cancer cell lines SKOV3/A were found in RGD-Lipo-siRNA preparation as compared to that of unmodified cationic lipsomes (Lipo-siRNA). Using the post-insertion method for RGD modification, lysosome release of siRNA in pRGD-Lipo-siRNA was improved. From flow cytometry, significant increase of doxorubicin accumulation was observed in the SKOV3/A cells treated with pRGD-Lipo-siRNA targeting human MDR1 gene. In vitro cytotoxicity assay showed that the significant cell growth inhibition was achieved in the SKOV3/A cells after treating with the combined use of siRNA and doxorubicin. In conclusions, postinserted RGD modified lipoplex, pRGD-Lipo-siRNA, was successfully used for siRNA transfection and achieved drug resistance reversal in human ovarian cancer SKOV3/A (doxorubicin-resistant) cells. It suggested that this liposomes might be a potential vehicle for siRNA delivery in vivo.

Key words: siRNA, siRNA, Cationic liposomes, Cationic liposomes, DC-Chol, DC-Chol, RGD modification, RGD modification, ,

中图分类号:

R943.42

Supporting: Foundation items: National Natural Science Foundation of China (Grant No. 30701056), Foundation of MOST (973 Program, Grant No. 2007CB935801), Beijing Natural Science Foundation of China (Grant No. 7083112) and Doctoral Foundation of Ministry of Education of China (Grant No. 20070001813).
^*Corresponding author. Tel./fax: 86-10-82802683;

杨世进, 杨丽娟, 姜娟, 王坚成^*, 张强.

整合素配体RGD修饰的阳离子脂质体作为siRNA输送载体的体外研究

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Shi-Jin Yang, Li-Juan Yang, Juan Jiang, Jian-Cheng Wang^*, Qiang Zhang. In vitro investigation of RGD-modified stabilized cationic liposomes as non-viral vehicle for siRNA delivery[J]. .

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