The long-term survival rate of cancer patients has been increasing as a result of advances in treatments and precise medical management. The evidence has accumulated that the incidence and mortality of non-cancer diseases have increased along with the increase in survival time and long-term survival rate of cancer patients after radiotherapy. The risk of cardiovascular disease as a radiation late effect of tissue damage reactions is becoming a critical challenge and attracts great concern. Epidemiological research and clinical trials have clearly shown the close association between the development of cardiovascular disease in long-term cancer survivors and radiation exposure. Experimental biological data also strongly supports the above statement. Cardiovascular diseases can occur decades post-irradiation, and from initiation and development to illness, there is a complicated process, including direct and indirect damage of endothelial cells by radiation, acute vasculitis with neutrophil invasion, endothelial dysfunction, altered permeability, tissue reactions, capillary-like network loss, and activation of coagulator mechanisms, fibrosis, and atherosclerosis. We summarize the most recent literature on the tissue reactions and mechanisms that contribute to the development of radiation-induced cardiovascular diseases (RICVD) and provide biological knowledge for building preventative strategies.

TAB182 participates in DNA damage repair and radio-/chemosensitivity regulation in various tumors, but its role in tumorigenesis and therapeutic resistance in breast cancer remains unclear. In the current paper, we observed that triple-negative Breast Cancer (TNBC), a highly aggressive type of breast cancer, exhibits a lower expression of TAB182. TAB182 knockdown stimulates the proliferation, migration, and invasion of TNBC cells. Our study first obtained RNA-seq data to explore the cellular functions mediated by TAB182 at the genome level in TNBC cells. A transcriptome analysis and in vitro experiments enabled us to identify that TAB182 downregulation drives the enhanced properties of cancer stem-like cells (CSCs) in TNBC cells. Furthermore, TAB182 deletion contributes to the resistance of cells to olaparib or cisplatin, which can be rescued by silencing GLI2, a gene downstream of cancer stemness-related signaling pathways. Our results reveal a novel function of TAB182 as a potential negative regulator of cancer stem-like properties and drug sensitivity in TNBC cells, suggesting that TAB182 may be a tumor suppressor gene and is associated with increased therapeutic benefits for TNBC patients.

Mitochondrion is an important organelle of eukaryotic cells and a critical target of ionizing radiation (IR) outside the nucleus. The biological significance and mechanism of the non-target effect originating from mitochondria have received much attention in the field of radiation biology and protection. In this study, we investigated the effect, role, and radioprotective significance of cytosolic mitochondrial DNA (mtDNA) and its associated cGAS signaling on hematopoietic injury induced by IR in vitro culture cells and in vivo total body irradiated mice in this study. The results demonstrated that gamma-ray exposure increases the release of mtDNA into the cytosol to activate cGAS signaling pathway, and the voltage-dependent anion channel (VDAC) may contribute to IR-induced mtDNA release. VDAC1 inhibitor DIDS and cGAS synthetase inhibitor can alleviate bone marrow injury and ameliorate hematopoietic suppression induced by IR via protecting hematopoietic stem cells and adjusting subtype distribution of bone marrow cells, such as attenuating the increase of the F4/80+ macrophage proportion in bone marrow cells. The present study provides a new mechanistic explanation for the radiation non-target effect and an alternative technical strategy for the prevention and treatment of hematopoietic acute radiation syndrome.

目的:矽尘暴露引起肺部EMT（上皮间质转化）变化继而导致发生肺纤维化已有研究报道,其机制有待阐明。有研究报道证实多个lncRNA通过竞争性内源性机制与miRNA调控下游EMT相关基因表达,另有研究发现p53在EMT过程中发生重要作用。我们推测p53在矽尘所致EMT中的作用可能与lncRNA和miRNA的功能密切相关。本研究的目的是探讨矽尘诱导EMT过程中,p53/lncRNA/miRNA发挥的作用及其机制。材料与方法:本研究从细胞、动物、人群三个层面展开研究,采用二种细胞株:HBE细胞株和BEAS2b细胞株;使用Lipofectamine 2000试剂盒进行转染,CCK-8试剂盒分析细胞增殖情况,免疫印迹分析相关蛋白表达,FITC Annexin V凋亡检测试剂盒检测细胞凋亡情况,CRISPR-cas技术、siRNA干扰分别敲除或敲低HBE细胞株基因p53的表达,RNA免疫共沉淀（RIP）方法分析RNA和蛋白结合情况,HE染色病理切片观察EMT变化,免疫组化观察EMT相关基因表达定位和半定量情况,双荧光素报告酶基因检测p53对lncRNA的调控情况,qRT-PCR方法分析RNA相对表达情况。免疫印迹分析相关蛋白表达。结果:（1）HBE细胞,BEAS2b细胞在矽尘暴露后,p53、lncRNA RMRP表达随着暴露时间的延长而升高,而miRNA122表达随着暴露时间的延长而降低。（2）沉默p53表达后,lncRNA RMRP表达下降,miRNA122表达升高;沉默lncRNA RMRP表达后,p53表达降低,miRNA122表达升高;细胞加入miRNA122 mimic后,p53、lncRNA RMRP表达下降;（3）分别敲除p53和lncRNA RMRP,EMT相关生物标志物如上皮细胞标志物E-cadherine表达随矽尘暴露时间的延长而不断升高,而间质标志物N-cadherin、Vimentin等随着矽尘暴露时间的延长不断表达降低,敲除miRNA122,E-cadherine表达随着矽尘暴露时间的延长而不断降低,而间质标志物N-cadherin、Vimentin等随着矽尘暴露时间的延长不断表达增加;（4）动物层面上,lncRNA RMRP过表达载体导入小鼠后,肺组织p53表达随矽尘暴露时间延长而不断表达升高,miRNA122表达与之相反,进一步将miRNA122mimic导入小鼠体内,肺组织p53表达随矽尘暴露时间延长而不断表达降低,同时EMT相关标志物的变化情况与细胞实验趋势一致;（5）双荧光酶报告基因分析发现p53与lncRNARMRP上游潜在p53结合位点结合,启动lncRNA RMRP的表达;miRNA 122直接与p53的3‘-UTR区域结合,降低p53的表达。RIP进一步证实p53能与lncRNA RMRP结合;（6）lncRNA RMRP通过激活Notch通路发挥促进EMT作用。结论:lncRNA RMRP在矽尘暴露后升高,促进EMT的进程。P53调控lncRNA RMRP的表达,使其表达升高,升高后的lncRNA RMRP能竞争性与miR122结合,降低MiR122的表达,使得miR122对下游靶基因p53解除抑制作用,p53表达升高,p53/lncRNA RMRP/miR122之间形成反馈回路,随着矽尘暴露时间延长,lncRNA RMRP对EMT的作用不断放大。p53/lncRNA RMRP/miR122回路在EMT进程中起到促进作用。

BACKGROUND: TAB182 is overexpressed in cancerous tissues and correlated with poor overall survival in lung cancer patients. Mechanistically, TAB182 participates in DNA damage repair and endows tumour cells with radio- and chemoresistance. However, its role in non-small cell lung cancer (NSCLC) remains unclear.; METHODS AND RESULTS: Cells with stable TAB182 knockdown (KD) were generated using A549 NSCLC cells, and we demonstrated that depleting TAB182 inhibits cell EMT, proliferation, colony formation, migration and invasion. Analysis of the TCGA database showed a positive correlation between TAB182 and EGFR, a well-established NSCLC oncoprotein. Then, we verified that silencing TAB182 decreases EGFR expression at both the mRNA and protein levels. Moreover, both TAB182 and EGFR were reported to restore ionizing radiation (IR)-triggered DNA damage. We validated that IR elevates the protein level of EGFR and that silencing TAB182 can alleviate IR-induced EGFR upregulation. Furthermore, overexpressing EGFR abrogates the inhibitory effects of TAB182 KD on EMT, migration, and invasion in A549 cells.; CONCLUSIONS: Our data demonstrated that EGFR expression is regulated by TAB182 and downregulation of TAB182 has a novel function to repress EMT, migration and invasion by decreasing EGFR, indicating TAB182 could regulate the malignant progression of NSCLC. © 2023. The Author(s), under exclusive licence to Springer Nature B.V.

DNA双链断裂（DSBs）是电离辐射诱发最严重的DNA损伤类型,在哺乳动物细胞中DSBs主要由非同源末端连接（NHEJ）与同源重组（HR）两个途径进行修复。核酸外切酶EXO1介导的DSBs末端单链剪切在HR修复以及ATR-CHK1途径诱导的G2/M周期阻滞中发挥着重要调控作用,但其具体机制还不清楚。ZGRF1被鉴定为参与DNA损伤修复的重要解旋酶,但对其具体的作用机制鲜有研究,ZGRF1参与损伤修复的分子过程有待揭示。目的:研究ZGRF1在电离辐射诱发DNA损伤修复中的作用,探索ZGRF1参与的DNA修复通路及调控点、与已知的关键DNA修复蛋白的相互作用和相互调节的关系,阐述其主要的生物学意义。方法:（1）通过质谱分析鉴定辐照后ZGRF1形成的复合体中的蛋白质组分,采用免疫共沉淀（Co-IP）验证辐照前后ZGRF1与复合体成分中BRCA1、EXO1等的相互作用;（2）免疫荧光共定位（IF）检验在电离辐射损伤的HeLa细胞中ZGRF1与DSBs分子标志物γH2AX的共定位情况,ZGRF1与BRCA1、53BP1、MDC1、RNF8、EXO1等修复蛋白在DNA双链断裂位点的共定位;（3）检测ZGRF1敲除的HeLa细胞中DNA断裂末端单链DNA（3’ss DNA）生成标志物——RPA2在双链断裂位点的焦点（foci）形成;（4）使用DR-GFP和EJ-GFP报告系统分析野生型和ZGRF1敲除的HeLa细胞的HR和NHEJ修复效率,确定ZGRF1参与的DNA修复途径;分别单独敲低BRCA1、EXO1、BLM或与ZGRF1双敲低,检测ZGRF1与上述分子在HR修复通路中的相互调节关系;（5）克隆形成实验检测野生型和ZGRF1敲除的HeLa细胞和MDA-MB-231细胞对辐照以及PARP抑制剂（PARPi）Olarparib的敏感性,选用SR50指标进一步评价细胞对PARPi的敏感性;（6）MTT实验分析ZGRF1敲除的HeLa细胞和MDA-MB-231细胞对其他DNA损伤诱导剂（ADR、Cisplatin、ETO、CPT、MMC）的敏感性;（7）免疫印迹检测辐照后RPA2、细胞周期调控蛋白CHK1、CHK2等蛋白表达水平及磷酸化水平;（8）流式细胞仪检测野生型与ZGRF1敲除细胞辐照后的周期分布和凋亡发生率,并对G2/M期细胞进行定量统计分析。用磷酸化组蛋白H3p Ser10抗体免疫染色标记有丝分裂期细胞,流式细胞仪检测辐照后野生型与ZGRF1敲除细胞中阳性细胞,并进行统计分析;用诺考达唑处理细胞,使细胞停滞在有丝分裂期,流式细胞仪检测辐照后野生型与ZGRF1敲除细胞中有丝分裂期细胞比例。结果:（1）通过质谱分析和免疫共沉淀分析,发现ZGRF1与DNA修复相关蛋白BRCA1和EXO1存在相互作用,并且在电离辐射诱导DNA损伤后相互作用增强;（2）电离辐射后,ZGRF1与γH2AX形成共定位聚焦点,在受照射2小时之后ZGRF1敲除细胞中残留的γH2AX焦点数量显著高于野生型细胞,表明ZGRF1敲除细胞的DNA双链断裂修复效率降低。进一步分析显示,敲除ZGRF1导致DNA同源重组修复效率降低40%。在ZGRF1敲除细胞中再分别敲低BRCA1、EXO1,细胞的同源重组修复效率与单独敲低BRCA1、EXO1无明显差异,但略比单敲ZGRF1的修复效率低,同时敲低ZGRF1和BLM后细胞的同源重组修复效率降低更加显著,且低于二者任一单独敲低细胞的修复效率,提示ZGRF1与BLM在HR修复通路中的解旋酶作用有一定的互补性;（3）ZGRF1向DSB位点的募集受到BRCA1、MDC1和RNF8的影响,并不受53BP1的影响。敲除ZGRF1,RPA2焦点的形成显著减少,表明ZGRF1参与对DSBs断裂末端的单链DNA剪切形成3’ss DNA的调控;（4）敲除ZGRF1,抑制了IR诱导的RPA2和CHK1的磷酸化,但对RPA2和CHK1的蛋白总水平没有影响;（5）野生型与ZGRF1敲除的HeLa细胞的G2和M期细胞所占的百分比在照射后早期无明显差异。在照射后4-8小时,G2和M期细胞明显上升,但ZGRF1敲除细胞组的G2和M期细胞的百分比显著低于野生型细胞,而在照射后12小时,ZGRF1敲除细胞的G2和M期细胞百分比恢复至野生型水平。进一步分析M期细胞比例的变化发现,野生型在照射后1小时有丝分裂期细胞数量即明显减少,ZGRF1敲除细胞在照射后2小时后才开始明显减少,照后4小时则与野生型组数量无差异,恢复至野生型水平,表明敲除ZGRF1导致受照射细胞周期G2/M阻滞延迟;（6）ZGRF1敲除后的HeLa细胞和MDA-MB-231细胞在电离辐射后凋亡率增加,对电离辐射、PARP抑制剂Olaparib等DNA损伤化疗药物的敏感性显著增加。结论:ZGRF1参与调节电离辐射诱发DNA双链断裂的同源重组修复通路活性,揭示其通过协同EXO1的作用,介导DNA双链断裂末端单链剪切形成3’ss DNA,从而促进HR修复和G2/M周期阻滞的新机制。ZGRF1依赖于BRCA1与MDC1-RNF1通路募集到DNA损伤位点,在HR修复通路上可能与BLM的解旋酶活性存在功能互补。此外,ZGRF1敲除增加宫颈癌和乳腺癌细胞系对PARPi、IR等多种DNA损伤剂处理的敏感性。该研究为电离辐射所致DNA损伤修复调控和细胞放射敏感性提供了新的机制阐述,也为临床肿瘤的治疗提供了新的潜在靶点。

转化生长因子β1(TGF-β1)是一种重要的成纤维细胞因子,其表达水平能反映肺纤维化的严重程度。放射性肺损伤(RILI)包括早期的放射性肺炎(RP)和晚期的放射性肺纤维化(RIPF)。RP向RIPF发展的过程中伴随着TGF-β1表达水平的升高。了解及掌

Background Ionizing radiation (IR) can induce pulmonary fibrosis by causing epithelial mesenchymal transition (EMT), but the exact mechanism has not been elucidated. To investigate the molecular mechanism of how radiation induces pulmonary fibrosis by altering miR-486-3p content and thus inducing EMT. Methods The changes of miR-486-3p in cells after irradiation were detected by RT-qPCR. Western blot was used to detect the changes of cellular epithelial marker protein E-cadherin, mesenchymal marker N-cadherin, Vimentin and other proteins. The target gene of miR-486-3p was predicted by bioinformatics method and the binding site was verified by dual luciferase reporter system. In vivo experiments, adeno-associated virus (AAV) was used to carry miR-486-3p mimic to lung. Radiation-induced pulmonary fibrosis (RIPF) model was constructed by 25Gy(60)Co gamma-rays. The structural changes of mouse lung were observed by HE and Masson staining. The expression of relevant proteins in mice was detected by immunohistochemistry. Results IR could decrease the miR-486-3p levels in vitro and in vivo, and that effect was closely correlated to the occurrence of RIPF. The expression of Snail, which induces EMT, was shown to be restrained by miR-486-3p. Therefore, knockdown of Snail blocked the EMT process induced by radiation or knockdown of miR-486-3p. In addition, the molecular mechanism underlying the IR-induced miRNA level reduction was explored. The increased in BCL6 could inhibit the formation of pri-miR-486-3p, thereby reducing the levels of miR-486-3p in the alveolar epithelial cells, which would otherwise promote EMT and contribute to RIPF by targeting Snail. Conclusion IR can exacerbate RIPF in mice by activating the transcription factor BCL6, which inhibits the transcription of miR-486-3p and decreases its content, which in turn increases the content of the target gene slug and triggers EMT.