p53 Promoted Ferroptosis in Ovarian Cancer Cells Treated with Human Serum Incubated-Superparamagnetic Iron Oxides.
p53 Promoted Ferroptosis in Ovarian Cancer Cells Treated with Human Serum Incubated-Superparamagnetic Iron Oxides.
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p53 促进人血清孵育超顺磁性氧化铁处理的卵巢癌细胞铁死亡
DOI：
10.2147/ijn.s282489
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发表时间：
2021
影响因子：
8
通讯作者：
Yu H
中科院分区：
文献类型：
--
作者：
Zhang Y;Xia M;Zhou Z;Hu X;Wang J;Zhang M;Li Y;Sun L;Chen F;Yu H
方法采用MTT法检测SPIO血清和野生型p53过表达联合应用对卵巢癌细胞的抑制作用。普鲁士蓝染色和铁测定用于确定SPIO-血清处理后细胞内铁浓度的变化。TEM用于评估SPIO-血清处理诱导的任何线粒体损伤，Western印迹用于评估铁转运蛋白和脂质过氧化调节蛋白的表达。JC-1用于测量线粒体膜电位，并通过流式细胞术估计ROS水平。最后，使用荧光显微镜确认xCT蛋白表达和线粒体ROS水平。结果SPIO血清能有效诱导脂质过氧化反应，产生大量毒性活性氧。它还促进GPX 4和xCT的下调，最终导致铁依赖性氧化死亡。铁螯合剂DFO和脂质过氧化抑制剂Fer-1可逆转上述作用。SPIO-血清处理通过调节铁摄取破坏细胞内铁稳态，并且细胞呈现缺失的线粒体嵴和破裂的线粒体外膜。此外，我们能够表明，p53有助于SPIO血清诱导的卵巢癌细胞中的铁凋亡。结论SPIO血清可诱导卵巢癌细胞铁凋亡，p53过表达可促进卵巢癌细胞铁凋亡。我们的研究结果为铁凋亡作为一种新的纳米材料诱导的细胞死亡表型提供了理论依据。
Methods In this study, we used MTT assays to demonstrate that a combination of SPIO-Serum and wild-type p53 overexpression can reduce ovarian cancer cell viability in vitro. Prussian blue staining and iron assays were used to determine changes in intracellular iron concentration following SPIO-Serum treatment. TEM was used to evaluate any mitochondrial damage induced by SPIO-Serum treatment, and Western blot was used to evaluate the expression of the iron transporter and lipid peroxidation regulator proteins. JC-1 was used to measure mitochondrial membrane potential, and ROS levels were estimated by flow cytometry. Finally, xCT protein expression and mitochondrial ROS levels were confirmed using fluorescence microscopy. Results SPIO-Serum effectively induced lipid peroxidation and generated abundant toxic ROS. It also facilitated the downregulation of GPX4 and xCT, ultimately resulting in iron-dependent oxidative death. These effects could be reversed by iron chelator DFO and lipid peroxidation inhibitor Fer-1. SPIO-Serum treatment disrupted intracellular iron homeostasis by regulating iron uptake and the cells presented with missing mitochondrial cristae and ruptured outer mitochondrial membranes. Moreover, we were able to show that p53 contributed to SPIO-Serum-induced ferroptosis in ovarian cancer cells. Conclusion SPIO-Serum induced ferroptosis and overexpressed p53 contributed to ferroptosis in ovarian cancer cells. Our data provide a theoretical basis for ferroptosis as a novel cell death phenotype induced by nanomaterials.