Intracellular uptake and trafficking of pluronic micelles in drug-sensitive and MDR cells: Effect on the intracellular drug localization
Intracellular uptake and trafficking of pluronic micelles in drug-sensitive and MDR cells: Effect on the intracellular drug localization
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DOI：
10.1002/jps.10006
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发表时间：
2002-01-01
影响因子：
3.8
通讯作者：
Muniruzzaman, M
中科院分区：
文献类型：
--
作者：
Rapoport, N;Marin, A;Muniruzzaman, M
用流式细胞术和荧光显微镜观察了荧光标记的Pluronic P-105在HL-60白血病细胞、A2780药物敏感性卵巢癌细胞和A2780/ADR MDR卵巢癌细胞中的摄取和定位。Pluronic P-105分子用pH敏感性荧光标记物5-（和6-）羧基-2 '7 '-二氯荧光素标记。标记的普朗尼克的荧光强度在pH 7.4下比在pH 5.5下高约两倍。在Pluronic浓度超过临界胶束浓度（CMC），流式细胞术直方图表现出双峰分布的细胞荧光的所有类型的细胞。具有较高荧光强度特征的细胞群体可能是由于Pluronic从细胞质囊泡（内体或溶酶体）的酸性环境转移到细胞质和细胞核的中性环境中，这表明Pluronic分子对酸性囊泡的膜进行了透化。对于MDR细胞，在孵育培养基中非常低的普朗尼克浓度下已经观察到细胞荧光的双峰分布（即，低于CMC）。这些数据表明，MDR细胞的酸性囊泡的膜比药物敏感细胞的酸性囊泡的膜更容易受到聚合物表面活性剂的作用。酸性囊泡的透化对药物的细胞内运输有显著影响：当在PBS中递送时，蒽环类药物ruboxyl（Rb）被隔离在细胞质囊泡中并被排除在细胞核之外;然而，当在Pluronic胶束中递送时，药物在细胞核中积累。药物从/与普朗尼克胶束的吸收，大大提高了超声。这些研究结果表明，通过液相内吞作用内化的药物的核积累可以通过应用普朗尼克胶束来增强，并且可以通过超声照射来进一步增强。(C)2002 Wiley-Liss，Inc.和美国制药协会。
The intracellular uptake and localization of a fluorescently labeled Pluronic P-105 in HL-60 leukemia cells and in A2780 drug-sensitive and A2780/ADR MDR ovarian carcinoma cells were characterized by flow cytometry and fluorescence microscopy. Pluronic P-105 molecules were labeled with a pH-sensitive fluorescent label, 5-(and 6-)carboxy-2'7'-dichlorofluorescein. The fluorescence intensity of labeled Pluronic was about twofold higher at pH 7.4 than at pH 5.5. At Pluronic concentrations exceeding the critical micelle concentration (cmc), flow cytometry histograms manifested bimodal distribution of cell fluorescence for all types of cells. Cell population characterized by higher fluorescence intensity presumably resulted from Pluronic transfer from the acidic environment of cytoplasmic vesicles (endosomes or lysosomes) into the neutral environment of the cytoplasm and cell nuclei, which suggested the permeabilization of the membranes of acidic vesicle by Pluronic molecules. For the MDR cells, the bimodal distribution of cell fluorescence was already observed at very low Pluronic concentrations in the incubation medium (i.e., below the cmc). The data suggest that the membranes of acidic vesicles of MDR cells are more susceptible to the action of polymeric surfactants than those of drug-sensitive cells. Permeabilization of acidic vesicles had a dramatic effect on the intracellular trafficking of drugs: when delivered in PBS, the anthracyclin drug ruboxyl (Rb) sequestered in cytoplasmic vesicles and was excluded from cell nuclei; however, when delivered in Pluronic micelles, drug accumulated in cell nuclei. Drug uptake from/with Pluronic micelles was substantially enhanced by ultrasound. These findings suggest that the nuclear accumulation of drugs internalized via fluid-phase endocytosis can be enhanced by the application of Pluronic micelles and can be further augmented by ultrasonic irradiation. (C) 2002 Wiley-Liss, Inc. and the American Pharmaceutical Association.