Effect of X-rays on transcript expression of rat brain microvascular endothelial cells: role of calcium signaling in X-ray-induced endothelium damage

Wu, Qibing; Fang, Yang; Zhang, Xinchen; Song, Fei; Wang, Yan; Chen, Hongbo; Du, Juan; Zheng, Chang‐Bo; Shen, Bing

doi:10.1042/bsr20193760

Cited by 9 publications

(11 citation statements)

References 42 publications

(60 reference statements)

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“…However, how the underlying mechanisms of endothelial dysfunction and endothelial function change over time and with dose of radiation remain unclear. We previously investigated the effect of a delivered x-ray dose of 20 Gy on transcript expression in BMECs by using high-throughput sequencing ( Wu et al, 2020 ), a powerful tool for revealing changed genetic patterns and potential therapeutic targets of disease ( Ma et al, 2010 ; Azorsa et al, 2016 ). The present study followed up on that work and further explored transcript expression alterations with two doses (5 Gy or 20 Gy) and times (12 h or 24 h) following x-ray exposure.…”

Section: Discussionmentioning

confidence: 99%

“…However, such data assessing the molecular mechanisms of radiation-induced vascular dysfunction are insufficient and incomplete. A recent study by our team analyzed x-ray–treated (20 Gy) brain microvascular endothelial cells (BMECs) with next-generation sequencing and found that this radiation causes 383 genes to be significantly differentially expressed in BMECs and that these genetic alterations are accompanied by functional changes in the calcium (Ca 2+ ) signaling pathway ( Wu et al, 2020 ). In the present study, we used next-generation sequencing to further investigate the effects of two delivered doses of ionizing radiation (5 and 20 Gy) and of time after the exposure on transcriptional expression changes in BMECs.…”

Section: Introductionmentioning

confidence: 99%

“…Because ionizing irradiation induces alterations in the Ca 2+ signaling pathway in BMECs ( Wu et al, 2020 ), it is necessary to determine the molecular mechanisms underlying the potential Ca 2+ signaling pathway–induced brain injury. The most widely known pathway for entry of Ca 2+ into cells is through store-operated Ca 2+ entry (SOCE), and SOCE is modulated by STIM, a Ca 2+ sensor located in the endoplasmic reticulum, and by proteins in the Orai family, which form highly selective Ca 2+ channels ( Moccia et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

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X-Ray Causes mRNA Transcripts Change to Enhance Orai2-Mediated Ca2+ Influx in Rat Brain Microvascular Endothelial Cells

Yang

Gao

et al. 2021

Front. Mol. Biosci.

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Background: Radiation-induced brain injury is a serious and treatment-limiting complication of brain radiation therapy. Although endothelial cell dysfunction plays a critical role in the development of this pathogenesis, the underlying molecular mechanisms remain elusive.Methods: Primary cultured rat brain microvascular endothelial cells (BMECs) were divided into five groups without or with exposure of x-rays delivered at 5 Gy or 20 Gy. For the irradiated groups, cells were continued to cultivate for 12 or 24 h after being irradiated. Then the mRNA libraries of each group were established and applied for next-generation sequencing. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were conducted to analyze the sequencing results. Quantitative polymerase chain reaction, western blotting, cck8 assay and intracellular calcium concentration assays were conducted to analyze the role of Orai2-associated SOCE in x-ray induced cellular injury.Results: In total, 3,005 transcripts in all the four x-ray–exposed groups of BMECs showed expression level changes compared with controls. With the dose of x-ray augment and the following cultured time extension, the numbers of differentially expressed genes (DEGs) increased significantly in BMECs. Venn diagrams identified 40 DEGs common to all four exposure groups. Functional pathway enrichment analyses indicated that those 40 DEGs were enriched in the calcium signaling pathway. Among those 40 DEGs, mRNA and protein expression levels of Orai2 were significantly upregulated for 24 h. Similarly, calcium influx via store-operated calcium entry, which is modulated by Orai2, was also significantly increased for 24 h in x-ray–exposed BMECs. Moreover, the change in SOCE was suppressed by btp-2, which is a non-selective inhibitor of Orai. Additionally, x-ray exposure induced a significant decrease of proliferation in BMECs in the dose- and time-dependent manner.Conclusion: These findings provide evidence for molecular mechanisms underlying BMECs dysfunction in development of radiation-induced brain injury and suggest new approaches for therapeutic targets.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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X-Ray Causes mRNA Transcripts Change to Enhance Orai2-Mediated Ca2+ Influx in Rat Brain Microvascular Endothelial Cells

Yang

Gao

et al. 2021

Front. Mol. Biosci.

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“…Cell death, proliferation and clonogenic ability are affected by irradiation depending on the dose delivered and the characteristics of the beam. Irradiation-related apoptosis was the most studied effect (43/147 studies, 29.3%), mainly measured through immunofluorescence staining ( Figure 4 a) or flow cytometry [ 13 , 16 , 23 , 30 , 32 , 46 , 61 , 63 , 66 , 68 , 72 , 73 , 74 , 75 , 76 , 77 , 78 , 79 , 80 , 81 , 82 , 83 , 84 , 85 , 86 , 87 , 88 , 89 , 90 , 91 , 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 , 100 , 101 , 102 , 103 , 104 , 105 ]. The causal link between IR and apoptosis in ECs has been investigated for years, with examples going back to 25 years ago [ 103 ].…”

Section: Analyzing Resultsmentioning

confidence: 99%

Radiobiological Studies of Microvascular Damage through In Vitro Models: A Methodological Perspective

Possenti

Mecchi

Rossoni

et al. 2021

Cancers

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Ionizing radiation (IR) is used in radiotherapy as a treatment to destroy cancer. Such treatment also affects other tissues, resulting in the so-called normal tissue complications. Endothelial cells (ECs) composing the microvasculature have essential roles in the microenvironment’s homeostasis (ME). Thus, detrimental effects induced by irradiation on ECs can influence both the tumor and healthy tissue. In-vitro models can be advantageous to study these phenomena. In this systematic review, we analyzed in-vitro models of ECs subjected to IR. We highlighted the critical issues involved in the production, irradiation, and analysis of such radiobiological in-vitro models to study microvascular endothelial cells damage. For each step, we analyzed common methodologies and critical points required to obtain a reliable model. We identified the generation of a 3D environment for model production and the inclusion of heterogeneous cell populations for a reliable ME recapitulation. Additionally, we highlighted how essential information on the irradiation scheme, crucial to correlate better observed in vitro effects to the clinical scenario, are often neglected in the analyzed studies, limiting the translation of achieved results.

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“…Our team recently used next-generation sequencing to explore changes in the transcription profile of rat brain microvascular endothelial cells (rBMECs) after radiation exposure [ 8 ]. We found that calcium (Ca 2+ ) signals including 38 genes were significantly changed.…”

Section: Introductionmentioning

confidence: 99%

Role of Orai3-Mediated Store-Operated Calcium Entry in Radiation-Induced Brain Microvascular Endothelial Cell Injury

Fang

Huang

et al. 2023

IJMS

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Radiation-induced brain injury is a serious complication with complex pathogenesis that may accompany radiotherapy of head and neck tumors. Although studies have shown that calcium (Ca2+) signaling may be involved in the occurrence and development of radiation-induced brain injury, the underlying molecular mechanisms are not well understood. In this study, we used real-time quantitative polymerase chain reaction and Western blotting assays to verify our previous finding using next-generation sequencing that the mRNA and protein expression levels of Orai3 in rat brain microvascular endothelial cells (rBMECs) increased after X-ray irradiation. We next explored the role of Orai3 and Orai3-mediated store-operated Ca2+ entry (SOCE) in radiation-induced brain injury. Primary cultured rBMECs derived from wild-type and Orai3 knockout (Orai3(−/−)) Sprague–Dawley rats were used for in vitro experiments. Orai3-mediated SOCE was significantly increased in rBMECs after X-ray irradiation. However, X-ray irradiation-induced SOCE increase was markedly reduced in Orai3 knockout rBMECs, and the percentage of BTP2 (a nonselective inhibitor of Orai channels)-inhibited SOCE was significantly decreased in Orai3 knockout rBMECs. Functional studies indicated that X-ray irradiation decreased rBMEC proliferation, migration, and tube formation (a model for assessing angiogenesis) but increased rBMEC apoptosis, all of which were ameliorated by BTP2. In addition, occurrences of all four functional deficits were suppressed in X-ray irradiation-exposed rBMECs derived from Orai3(−/−) rats. Cerebrovascular damage caused by whole-brain X-ray irradiation was much less in Orai3(−/−) rats than in wild-type rats. These findings provide evidence that Orai3-mediated SOCE plays an important role in radiation-induced rBMEC damage and brain injury and suggest that Orai3 may warrant development as a potential therapeutic target for reducing or preventing radiation-induced brain injury.

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Effect of X-rays on transcript expression of rat brain microvascular endothelial cells: role of calcium signaling in X-ray-induced endothelium damage

Cited by 9 publications

References 42 publications

X-Ray Causes mRNA Transcripts Change to Enhance Orai2-Mediated Ca2+ Influx in Rat Brain Microvascular Endothelial Cells

X-Ray Causes mRNA Transcripts Change to Enhance Orai2-Mediated Ca2+ Influx in Rat Brain Microvascular Endothelial Cells

Radiobiological Studies of Microvascular Damage through In Vitro Models: A Methodological Perspective

Role of Orai3-Mediated Store-Operated Calcium Entry in Radiation-Induced Brain Microvascular Endothelial Cell Injury

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