HIV-TAT mediated protein transduction of Cu/Zn-superoxide dismutase-1 (SOD1) protects skin cells from ionizing radiation.
HIV-TAT mediated protein transduction of Cu/Zn-superoxide dismutase-1 (SOD1) protects skin cells from ionizing radiation.
复制标题
HIV-TAT 介导的铜/锌超氧化物歧化酶-1 (SOD1) 蛋白转导可保护皮肤细胞免受电离辐射。
DOI：
10.1186/1748-717x-8-253
复制
发表时间：
2013-10-31
期刊：
影响因子：
--
通讯作者：
Cao J
中科院分区：
文献类型：
--
作者：
Gu Q;Feng T;Cao H;Tang Y;Ge X;Luo J;Xue J;Wu J;Yang H;Zhang S;Cao J
放射治疗期间，放射性皮肤损伤仍然是一个严重的问题。铜/锌超氧化物歧化酶是一种保守的清除细胞内超氧阴离子自由基的酶。由于细胞膜的完整性，外源分子不能被掺入细胞内，限制了天然SOD1的应用。本研究的目的是评价HIV-TAT蛋白转导结构域介导的SOD1蛋白转导(TAT-SOD1)对电离辐射的保护作用。利用原核表达系统获得重组TAT-SOD1和SOD1。通过免疫荧光和抗氧化能力检测TAT-SOD1在人角质形成细胞HaCaT中的转导作用和生物学活性。采用Mito-Tracker染色、活性氧(ROS)生成试验、细胞凋亡分析和丙二醛(MDA)测定评价TAT-SOD1的保护作用。HaCaT细胞对TAT-SOD1的摄取保持了其生物学活性。与天然SOD1相比，应用TAT-SOD1可显著提高X射线照射诱导的细胞存活率，减少细胞凋亡率。此外，TAT-SOD1降低了辐射后HaCaT细胞的ROS，并保持了线粒体的完整性。以γ双链断裂为代表的辐射诱导的DNAH_2AX焦点经TAT-SOD处理后明显减少。此外，皮下注射TAT-SOD可显著降低45Gy射线诱发的大鼠皮肤ROS和MDA浓度。本研究为TAT-SOD1减轻HaCaT细胞和大鼠皮肤辐射损伤提供了证据，为放射性皮肤损伤的治疗提供了新的策略。
Radiation-induced skin injury remains a serious concern during radiotherapy. Cu/Zn-superoxide dismutase (Cu/Zn-SOD, SOD1) is a conserved enzyme for scavenging superoxide radical in cells. Because of the integrity of cell membranes, exogenous molecule is not able to be incorporated into cells, which limited the application of natural SOD1. The aim of this study was to evaluate the protective role of HIV-TAT protein transduction domain mediated protein transduction of SOD1 (TAT-SOD1) against ionizing radiation. The recombinant TAT-SOD1 and SOD1 were obtained by prokaryotic–based protein expression system. The transduction effect and biological activity of TAT-SOD1 was measured by immunofluorescence and antioxidant capability assays in human keratinocyte HaCaT cells. Mito-Tracker staining, reactive oxygen species (ROS) generation assay, cell apoptosis analysis and malondialdehyde (MDA) assay were used to access the protective effect of TAT- SOD1. Uptake of TAT-SOD1 by HaCaT cells retained its biological activity. Compared with natural SOD1, the application of TAT-SOD1 significantly enhanced the viability and decreased the apoptosis induced by X-ray irradiation. Moreover, TAT-SOD1 reduced ROS and preserved mitochondrial integrity after radiation exposure in HaCaT cells. Radiation-induced γH2AX foci, which are representative of DNA double strand breaks, were decreased by pretreatment with TAT-SOD1. Furthermore, subcutaneous application of TAT-SOD1 resulted in a significant decrease in 45 Gy electron beam-induced ROS and MDA concentration in the skins of rats. This study provides evidences for the protective role of TAT-SOD1 in alleviating radiation-induced damage in HaCaT cells and rat skins, which suggests a new therapeutic strategy for radiation-induced skin injury.