The Natural Cell-Penetrating Peptide Crotamine Targets Tumor Tissue in Vivo and Triggers a Lethal Calcium-Dependent Pathway in Cultured Cells
The Natural Cell-Penetrating Peptide Crotamine Targets Tumor Tissue in Vivo and Triggers a Lethal Calcium-Dependent Pathway in Cultured Cells
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DOI：
10.1021/mp2000605
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发表时间：
2012-02-01
影响因子：
4.9
通讯作者：
Hayashi, Mirian A. F.
中科院分区：
文献类型：
--
作者：
Nascimento, Fabio D.;Sancey, Lucie;Hayashi, Mirian A. F.
我们的目标是证明克罗胺在体内肿瘤特异性积累，克罗胺是一种来自南美响尾蛇Crotalus durissus terrificus毒液的天然肽，我们小组将其描述为一种细胞穿透肽，对活跃增殖的细胞具有高度特异性，并具有浓度依赖性细胞毒作用。克罗胺细胞毒性已被证明依赖于溶酶体的破坏和随后细胞内蛋白酶的激活。在这项工作中，我们通过共聚焦显微镜实时观察到，克罗胺的细胞毒性作用还涉及细胞内钙的快速释放和线粒体膜电位的损失。克罗他胺引起的细胞内钙超载几乎被毒胡萝卜素完全阻断。微荧光测定证实了内部细胞器（例如溶酶体和内质网）以及细胞外介质作为细胞内钙增加的贡献者的重要性。最后，我们在这里证明，腹腔注射的克罗胺可以有效地靶向移植在裸鼠体内的远端皮下肿瘤，正如非侵入性光学成像程序所证明的那样，该过程允许体内实时监测肿瘤组织中克罗胺的摄取。综上所述，我们的数据表明细胞毒性肽克罗胺可潜在用于双重目的：靶向和检测正在生长的肿瘤组织并选择性触发肿瘤细胞死亡。
Our goal was to demonstrate the in vivo tumor specific accumulation of crotamine, a natural peptide from the venom of the South American rattlesnake Crotalus durissus terrificus, which has been characterized by our group as a cell penetrating peptide with a high specificity for actively proliferating cells and with a concentration-dependent cytotoxic effect. Crotamine cytotoxicity has been shown to be dependent on the disruption of lysosomes and subsequent activation of intracellular proteases. In this work, we show that the cytotoxic effect of crotamine also involves rapid intracellular calcium release and loss of mitochondrial membrane potential as observed in real time by confocal microscopy. The intracellular calcium overload induced by crotamine was almost completely blocked by thapsigargin. Microfluorimetry assays confirmed the importance of internal organelles, such as lysosomes and the endoplasmic reticulum, as contributors for the intracellular calcium increase, as well as the extracellular medium. Finally, we demonstrate here that crotamine injected intraperitoneally can efficiently target remote subcutaneous tumors engrafted in nude mice, as demonstrated by a noninvasive optical imaging procedure that permits in vivo real-time monitoring of crotamine uptake into tumor tissue. Taken together, our data indicate that the cytotoxic peptide crotamine can be used potentially for a dual purpose: to target and detect growing tumor tissues and to selectively trigger tumor cell death.