NKCT1 (purified Naja kaouthia protein toxin) conjugated gold nanoparticles induced Akt/mTOR inactivation mediated autophagic and caspase 3 activated apoptotic cell death in leukemic cell
NKCT1 (purified Naja kaouthia protein toxin) conjugated gold nanoparticles induced Akt/mTOR inactivation mediated autophagic and caspase 3 activated apoptotic cell death in leukemic cell
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DOI：
10.1016/j.toxicon.2016.08.004
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发表时间：
2016-10-01
期刊：
影响因子：
2.8
通讯作者：
Gomes, Antony
中科院分区：
文献类型：
--
作者：
Bhowmik, Tanmoy;Gomes, Antony
背景：如前所述，金纳米颗粒(GNP)和蛇毒蛋白毒素NKCT1被偶联(Bhowmik等人，2013年)。本研究旨在探讨标记蛇毒蛋白毒素NKCT1的金纳米颗粒(GNP-NKCT1)对人白血病细胞株U937和K562的caspase依赖的凋亡途径和自噬诱导能力。方法：通过线粒体膜电位测定、ROS生成试验、caspase3途径和Western blotting检测GNP-NKCT1诱导U937和K562细胞凋亡的能力。Western blotting检测Akt、mTOR和PI3K的表达，检测GNP-NKCT1诱导的自噬途径。加入半胱氨酸天冬氨酸氨基转移酶3(Caspase3)抑制剂后，自噬细胞死亡情况也得到证实。结果：GNP-NKCT1诱导白血病细胞系后，线粒体膜电位下降，同时也表现出高ROS生成。细胞半胱氨酸天冬氨酸氨基转移酶3表达增加。在caspase3抑制剂作用下，GNP-NKCT1下调PI3K/Akt和mTOR的表达，从而导致自噬细胞死亡。溶酶体染色法证实溶酶体酶参与自噬反应。免疫印迹结果显示ATg3、Atg12、Beclin 1、Lc3-II蛋白和BIF-1表达上调，Atg4B表达下调。结论：金纳米颗粒与NKCT1偶联可诱导白血病细胞自噬以外的另一种细胞死亡途径。一般意义：本研究可能为从蛇毒等自然资源中开发化疗药物提供了解领域。(C)爱思唯尔有限公司出版的2016年。
Background: Gold nanoparticle (GNP) and snake venom protein toxin NKCT1 was conjugated as stated earlier (Bhowmik et al., 2013). The aim of this study was to explore the caspase dependent apoptotic pathway and autophagy inducing ability of gold nanoparticles tagged snake venom protein toxin NKCT1 (GNP-NKCT1) in human leukemic U937 and K562 cell line.Methods: GNP-NKCT1 induced apoptosis in U937 and 1(562 cell line were assessed through mitochondrial membrane potential assay, ROS generation assay, caspase 3 pathways and western blotting. GNP-NKCT1 induced autophagic pathway was detected through Akt, mTOR and PI3K expression by western blotting. Autophagic cell death also checked after addition of caspase 3 inhibitor and which also reconfirmed by western blotting of autophagic marker protein, lysosomal staining.Results: Loss of mitochondrial membrane potential was occurred in both the leukemic cell line after induction by GNP-NKCT1 and treatment of which also exhibited high ROS generation. Caspase 3 expression of cell was also increased. With caspase 3 inhibitor, GNP-NKCT1 downregulated PI3K/Akt and mTOR expression and thus undergoing autophagic cell death. Lysosomal staining confirmed lysosomal enzyme involvement in the autophagic response. Up regulation of Atg 3, Atg12, Beclin 1, LC3-II protein and BIF-1 and down regulation of Atg4B were also showed by blotting.Conclusion: The results demonstrated that conjugation of Gold nanoparticles with NKCT1 could induce an alternate cell death pathway other than apoptosis in the form of autophagy in leukemic cell. General significance: This study might provide the understanding area of chemotherapeutic drug development from natural resources like snake venoms. (C) 2016 Published by Elsevier Ltd.