State Key Lab of Radiation Medicine and Protection, Collaborative Innovation Center of Radiation Medicine of Jiangsu Higher Education Institutions, Soochow University, Suzhou, China.
Second Affiliated Hospital of Chengdu Medical College, China National Nuclear Corporation 416 Hospital, Chengdu, China.
J Dermatol Sci. 2020 Nov;100(2):139-147. doi: 10.1016/j.jdermsci.2020.09.003. Epub 2020 Sep 24.
Radiation-induced skin injury is one of the main adverse effects and a dose-limiting factor of radiotherapy without feasible treatment. The underlying mechanism of this disease is still limited.
To investigate the potential molecular pathways and mechanisms of radiation-induced skin injury.
mRNA expression profiles were determined by Affymetrix Human HTA2.0 microarray.IFI6 overexpression and knockdown were mediated by lentivirus. The functional changes of skin cells were measured by flow cytometry, ROS probe and Edu probe. Protein distribution was detected by immunofluorescence experiment, and IFI6-interacting proteins were detected by immunoprecipitation (IP) combined with mass spectrometry. The global gene changes in IFI6-overexpressed skin cells after irradiation were detected by RNA-seq.
mRNA expression profiling showed 50 upregulated and 13 down regulated genes and interferon alpha inducible protein 6 (IFI6) was top upregulated. Overexpression of IFI6 promoted cell proliferation and reduced cell apoptosis as well as ROS production following radiation, and conversely, increased the radiosensitivity of HaCaT and human skin fibroblast (WS1). IFI6 was translocated into nucleus in irradiated skin cells and the interacting relationship with mitochondrial single-stranded DNA-binding protein 1 (SSBP1), which could enhance the transcriptional activity of heat shock transcription factor 1 (HSF1).IFI6 augmented HSF1 activity following radiation in HaCaT and WS1 cells. RNA-seq analysis showed IFI6 modulated virus infection and cellular response to stress pathways, which may help to further explore how IFI6 regulate the transcriptional activity of HSF1.
This study reveals that IFI6 is induced by ionizing radiation and confers radioprotection in skin cells.
放射性皮肤损伤是放射治疗的主要不良反应之一,也是放射治疗的剂量限制因素,目前尚无可行的治疗方法。其发病机制尚不清楚。
探讨放射性皮肤损伤的潜在分子途径和机制。
采用 Affymetrix Human HTA2.0 微阵列检测 mRNA 表达谱。通过慢病毒转染过表达和敲低 IFI6。通过流式细胞术、ROS 探针和 Edu 探针检测皮肤细胞的功能变化。通过免疫荧光实验检测蛋白分布,通过免疫沉淀(IP)联合质谱检测 IFI6 相互作用蛋白。通过 RNA-seq 检测 IFI6 过表达的皮肤细胞在照射后的全基因变化。
mRNA 表达谱分析显示,50 个基因上调,13 个基因下调,干扰素α诱导蛋白 6(IFI6)上调最显著。IFI6 过表达可促进细胞增殖,减少细胞凋亡和 ROS 产生,同时降低 HaCaT 和人皮肤成纤维细胞(WS1)的放射敏感性。IFI6 在照射后的皮肤细胞中易位到核内,并与线粒体单链 DNA 结合蛋白 1(SSBP1)相互作用,从而增强热休克转录因子 1(HSF1)的转录活性。IFI6 增强了 HaCaT 和 WS1 细胞中 HSF1 的辐射后活性。RNA-seq 分析显示,IFI6 调节病毒感染和细胞对应激反应途径,这可能有助于进一步探讨 IFI6 如何调节 HSF1 的转录活性。
本研究揭示了 IFI6 是由电离辐射诱导的,并在皮肤细胞中赋予辐射保护作用。