MACROPHAGES LOADED WITH DOXORUBICIN BY ATP-MEDIATED PERMEABILIZATION - POTENTIAL CARRIERS FOR ANTITUMOR THERAPY
MACROPHAGES LOADED WITH DOXORUBICIN BY ATP-MEDIATED PERMEABILIZATION - POTENTIAL CARRIERS FOR ANTITUMOR THERAPY
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DOI：
10.1016/0167-4889(94)90200-3
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发表时间：
1994-11-10
影响因子：
5.1
通讯作者：
NASTRUZZI, C
中科院分区：
文献类型：
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作者：
MUNERATI, M;CORTESI, R;NASTRUZZI, C
在许多细胞类型中，细胞外 ATP (ATP(e)) 已被证明可引起低分子量 (< 900 Da) 水溶性化合物的可逆质膜透化。在本报告中，我们利用这项技术将分子质量为 543 Da 的抗癌药物阿霉素 (DXR) 掺入两种小鼠细胞系的细胞质中，这两种细胞系先前已被证明可表达 ATP(e) 门控孔、J774 巨噬细胞和肿瘤坏死因子 (TNF) 抗性 L929 成纤维细胞。与被动加载的细胞相比，ATP(e) 介导的可逆透化作用使 DXR 细胞内捕获增加至少 Lt 倍（0.5 pg/细胞与 2 pg/细胞）。 37 摄氏度下的释放动力学分析表明，在第一个小时内，约 40% 的细胞内 DXR 从 ATP(e) 透化细胞和被动负载细胞中释放出来；孵育 4 小时后观察到约 15% 的进一步释放。 DXR 释放曲线在 ATP(e) 透化细胞和被动负载细胞中相似。 ATP(e) 通透的、负载 DXR 的 (ATP(e)-DXR) 细胞强烈抑制 K562 肿瘤细胞的增殖。综上所述，这些结果表明 ATP(e) 介导的可逆质膜透化可以有效地用于用高浓度 DXR 负载不同组织型的细胞。这种方法可以通过使用活细胞来输送高剂量的抗癌药物，同时减少全身毒性作用。
In many cell types extracellular ATP (ATP(e)) has been shown to cause reversible plasma membrane permeabilization to low molecular weight (< 900 Da) water-soluble compounds. In the present report we have exploited this technique to incorporate the anticancer drug doxorubicin (DXR), molecular mass 543 Da, into the cytoplasm of two mouse cell lines that had previously been shown to express the ATP(e)-gated pore, J774 macrophages and tumor necrosis factor (TNF)-resistant L929 fibroblasts. Compared to passively loaded cells, ATP(e)-mediated reversible permeabilization allowed an at least Lt-fold increase in DXR intracellular trapping (0.5 pg/cell versus 2 pg/cell). Analysis of the release kinetics at 37 degrees C showed that about 40% of total intracellular DXR was discharged during the first hour from both ATP(e)-permeabilized and passively loaded cells; about 15% further release was observed upon incubation up to 4 h. DXR release profiles were similar in ATP(e)-permeabilized and passively loaded cells. ATP(e)-permeabilized, DXR-loaded (ATP(e)-DXR) cells strongly inhibited the proliferation of K562 tumor cells. Taken together these results indicate that ATP(e)-mediated reversible plasma membrane permeabilization can be effectively used to load cells of different histotypes with high concentrations of DXR. This approach could permit to vehicle high doses of anticancer agents by using living cells while reducing systemic toxic effects.