TAT peptide on the surface of liposomes affords their efficient intracellular delivery even at low temperature and in the presence of metabolic inhibitors
TAT peptide on the surface of liposomes affords their efficient intracellular delivery even at low temperature and in the presence of metabolic inhibitors
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DOI：
10.1073/pnas.151247498
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发表时间：
2001-07-17
影响因子：
11.1
通讯作者：
Levchenko, TS
中科院分区：
文献类型：
--
作者：
Torchilin, VP;Rammohan, R;Levchenko, TS
为了实现有效的细胞内药物和 DNA 递送，人们尝试绕过内吞途径将微粒药物载体靶向进入细胞质。源自 HIV-1 TAT 蛋白的 TAT 肽有助于蛋白质和小胶体颗粒的细胞内递送。我们证明，相对较大的药物载体，例如 200 nm 脂质体，也可以通过附着在脂质体表面的 TAT 肽递送到细胞中。脂质体用亲膜罗丹明-磷脂酰乙醇胺或通过包埋 FITC-葡聚糖进行荧光标记。将荧光 TAT 脂质体与小鼠 Lewis 肺癌细胞、人乳腺肿瘤 BT20 细胞和大鼠心肌细胞 H9C2 一起孵育，会导致某些脂质体在细胞内定位。 TAT 肽-细胞相互作用的空间位阻（TAT 直接附着到脂质体表面，无需间隔物或脂质体表面存在高分子量聚乙二醇）消除了脂质体内化，证明了 TAT 肽与细胞表面直接接触的重要性。低温或代谢抑制剂、叠氮化钠或碘乙酰胺对 TAT 脂质体转位到细胞内的影响很小，证实了该过程的能量独立特征。该方法可能对将药物直接输送到细胞质中具有重要意义。
To achieve an efficient intracellular drug and DNA delivery, attempts were made to target microparticulate drug carriers into cytoplasm bypassing the endocytotic pathway. TAT peptides derived from the HIV-1 TAT protein facilitate intracellular delivery of proteins and small colloidal particles. We demonstrated that relatively large drug carriers, such as 200-nm liposomes, can also be delivered into cells by TAT peptide attached to the liposome surface. Liposomes were fluorescently labeled with membranotropic rhodamine-phosphatidylethanolamine or by entrapping FITC-dextran. Incubation of fluorescent TAT liposomes with mouse Lewis lung carcinoma cells, human breast tumor BT20 cells, and rat cardiac myocyte H9C2 results in intracellular localization of certain liposomes. Steric hindrances for TAT peptide-cell interaction (attachment of TAT directly to the liposome surface without spacer or the presence of a high MW polyethylene glycol on the liposome surface) abolish liposome internalization, evidencing the importance of direct contact of TAT peptide with the cell surface. Low temperature or metabolic inhibitors, sodium azide or iodoacetamide, have little influence on the translocation of TAT liposomes into cells, confirming the energy-independent character of this process. The approach may have important implications for drug delivery directly into cell cytoplasm.