Radiation induces NORAD expression to promote ESCC radiotherapy resistance via EEPD1/ATR/Chk1 signalling and by inhibiting pri-miR-199a1 processing and the exosomal transfer of miR-199a-5p.
Radiation induces NORAD expression to promote ESCC radiotherapy resistance via EEPD1/ATR/Chk1 signalling and by inhibiting pri-miR-199a1 processing and the exosomal transfer of miR-199a-5p.
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辐射通过 EEPD1/ATR/Chk1 信号传导以及抑制 pri-miR-199a1 加工和 miR-199a-5p 的外泌体转移诱导 NORAD 表达，从而促进 ESCC 放疗抵抗
DOI：
10.1186/s13046-021-02084-5
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发表时间：
2021-09-29
期刊：
影响因子：
--
通讯作者：
Zhang X
中科院分区：
文献类型：
--
作者：
Sun Y;Wang J;Ma Y;Li J;Sun X;Zhao X;Shi X;Hu Y;Qu F;Zhang X
背景放射抵抗是一个知之甚少的现象，它导致放疗失败和随后的局部复发，威胁着很大一部分食管鳞癌患者。方法采用荧光原位杂交（FISH）和定量RT-PCR方法检测lncRNA-NORAD、pri-miR-199 a 1和miR-199 a-5 p在人乳腺癌细胞中的表达和定位。通过电子显微镜和纳米颗粒跟踪分析（NTA）观察和鉴定外泌体。进行高通量microRNA测序和TMT质谱以鉴定功能性miRNA和蛋白质。通过一系列的体内外实验研究了NORAD的生物学效应。ChIP，RIP-qPCR，co-IP和双荧光素酶报告基因检测进行了探索相关的RNA和protein.ResultsWe在这里显示，DNA损伤激活非编码RNA NORAD，这是ESCC辐射抗性的关键。NORAD在辐射抗性ESCC细胞中以高水平表达。辐射处理通过增强H3 K4 me 2在其序列中的富集来促进NORAD表达。NORAD敲除细胞在体内和体外对辐射表现出显著的超敏反应。北美防空司令部需要启动修复和重新启动失速叉，G2周期逮捕和同源重组修复后，辐射治疗。NORAD通过与pri-miR-199 a1竞争性结合miR-199 a1，抑制pri-miR-199 a1的加工，从而抑制miR-199 a-5 p的表达。NORAD敲除细胞中的成熟miR-199 a-5 p被包装到外泌体中; miR-199 a-5 p通过靶向EEPD 1然后抑制ATR/Chk 1信号通路来恢复辐射抗性细胞的辐射敏感性。同时，NORAD敲低抑制PD-L1的泛素化，从而更好地响应辐射和抗PD-1治疗在小鼠model.ConclusionsBased的研究结果，lncRNA-NORAD代表一个潜在的治疗目标，提高效率的免疫治疗结合放射在食管鳞癌。
BackgroundRadioresistance, a poorly understood phenomenon, results in the failure of radiotherapy and subsequent local recurrence, threatening a large proportion of patients with ESCC. To date, lncRNAs have been reported to be involved in diverse biological processes, including radioresistance.MethodsFISH and qRT–PCR were adopted to examine the expression and localization of lncRNA-NORAD, pri-miR-199a1 and miR-199a-5p. Electron microscopy and nanoparticle tracking analysis (NTA) were conducted to observe and identify exosomes. High-throughput microRNAs sequencing and TMT mass spectrometry were performed to identify the functional miRNA and proteins. A series of in vitro and in vivo experiments were performed to investigate the biological effect of NORAD. ChIP, RIP-qPCR, co-IP and dual-luciferase reporter assays were conducted to explore the interaction of related RNAs and proteins.ResultsWe show here that DNA damage activates the noncoding RNA NORAD, which is critical for ESCC radioresistance. NORAD was expressed at high levels in radioresistant ESCC cells. Radiation treatment promotes NORAD expression by enhancing H3K4me2 enrichment in its sequence. NORAD knockdown cells exhibit significant hypersensitivity to radiation in vivoandin vitro. NORAD is required to initiate the repair and restart of stalled forks, G2 cycle arrest and homologous recombination repair upon radiation treatment. Mechanistically, NORAD inhibits miR-199a-5p expression by competitively binding PUM1 from pri-miR-199a1, inhibiting the processing of pri-miR-199a1. Mature miR-199a-5p in NORAD knockdown cells is packaged into exosomes; miR-199a-5p restores the radiosensitivity of radioresistant cells by targeting EEPD1 and then inhibiting the ATR/Chk1 signalling pathway. Simultaneously, NORAD knockdown inhibits the ubiquitination of PD-L1, leading to a better response to radiation and anti-PD-1 treatment in a mouse model.ConclusionsBased on the findings of this study, lncRNA-NORAD represents a potential treatment target for improving the efficiency of immunotherapy in combination with radiation in ESCC.
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DOI：
10.1007/s10388-018-0641-9
发表时间：
2019-01
期刊：
Esophagus : official journal of the Japan Esophageal Society
影响因子：
--
作者：
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DOI：
10.1016/j.radonc.2016.06.017
发表时间：
2016-12
期刊：
Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology
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International Radiogenomics Consortium (RgC)
影响因子：
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16.8
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