Jiang Ge, Park Kitae, Kim Jiseok, Kim Ki Su, Oh Eun Ju, Kang Hyungu, Han Su-Eun, Oh Yu-Kyoung, Park Tae Gwan, Kwang Hahn Sei
Department of Material Science and Engineering, Pohang University of Science and Technology (POSTECH), San 31, Hyoja-dong, Nam-gu, Pohang, 790-784, Korea.
Biopolymers. 2008 Jul;89(7):635-42. doi: 10.1002/bip.20978.
A novel target specific small interfering RNA (siRNA) delivery system was successfully developed using polyethyleneimine (PEI)-hyaluronic acid (HA) conjugate. Anti-PGL3-Luc siRNA was used as a model system suppressing the PGL3-Luc gene expression. The siRNA/PEI-HA complex with an average size of ca. 21 nm appeared to be formed by electrostatic interaction between the negatively charged siRNA and the positively charged PEI of PEI-HA conjugate. The cytotoxicity of siRNA/PEI-HA complex to B16F1 cells was lower than that of siRNA/PEI complex according to the MTT assay. When B16F1 and HEK-293 cells were treated with fluorescein isothiocyanate (FITC) labeled siRNA/PEI-HA complex, B16F1 cells, with a lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), showed higher green fluorescent intensity than HEK-293 cells because of the HA receptor mediated endocytosis of the complex. Accordingly, the PGL3-Luc gene silencing of anti-PGL3-Luc siRNA/PEI-HA complex was more efficient in B16F1 cells than in HEK-293 cells. In addition, the inhibited PGL3-Luc gene silencing effect in the presence of free HA in the transfection medium revealed that siRNA/HA-PEI complex was selectively taken up to B16F1 cells via HA receptor mediated endocytosis. All these results demonstrated that the intracellular delivery of anti-PGL3-Luc siRNA/PEI-HA complex could be facilitated by the HA receptor mediated endocytosis.
利用聚乙烯亚胺（PEI）-透明质酸（HA）共轭物成功开发了一种新型的靶向特异性小干扰RNA（siRNA）递送系统。抗PGL3-Luc siRNA被用作抑制PGL3-Luc基因表达的模型系统。平均大小约为21 nm的siRNA/PEI-HA复合物似乎是由带负电荷的siRNA与PEI-HA共轭物带正电荷的PEI之间的静电相互作用形成的。根据MTT分析，siRNA/PEI-HA复合物对B16F1细胞的细胞毒性低于siRNA/PEI复合物。当用异硫氰酸荧光素（FITC）标记的siRNA/PEI-HA复合物处理B16F1和HEK-293细胞时，由于复合物通过HA受体介导的内吞作用，具有淋巴管内皮透明质酸受体-1（LYVE-1）的B16F1细胞显示出比HEK-293细胞更高的绿色荧光强度。因此，抗PGL3-Luc siRNA/PEI-HA复合物在B16F1细胞中的PGL3-Luc基因沉默比在HEK-293细胞中更有效。此外，转染培养基中存在游离HA时PGL3-Luc基因沉默作用受到抑制，这表明siRNA/HA-PEI复合物通过HA受体介导的内吞作用被选择性地摄取到B16F1细胞中。所有这些结果表明，HA受体介导的内吞作用可促进抗PGL3-Luc siRNA/PEI-HA复合物的细胞内递送。
