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环状 RNA_014511 通过与 miR-29b-2-5p 结合来影响骨髓间充质干细胞的放射敏感性。

CircRNA_014511 affects the radiosensitivity of bone marrow mesenchymal stem cells by binding to miR-29b-2-5p.

机构信息

Department of Clinical Laboratory, The Third Xiangya Hospital, Central South University, Hunan, China.

出版信息

Bosn J Basic Med Sci. 2019 May 20;19(2):155-163. doi: 10.17305/bjbms.2019.3935.

Abstract

Hematopoietic stem cell transplantation is commonly used in patients with certain hematological or bone marrow tumors. Total body irradiation combined with chemotherapy is part of the preconditioning protocol that was the most commonly used before hematopoietic stem cell transplantation. However, total body irradiation preconditioning damages other normal cells in bone marrow. Therefore, exploring the mechanism of radiation resistance in bone marrow mesenchymal stem cells is of great significance for recovering the hematopoietic function after cell transplantation. This study aimed to demonstrate the miR-29b adsorption of circRNA_014511 and explore the effect of circRNA_014511 on radiosensitivity of bone marrow mesenchymal stem cells. In this study, circRNA_014511 overexpression vector was constructed and transfected into bone marrow mesenchymal stem cells, miR-29b-2-5p and P53 were found to be decreased, which could be reversed by miR29b-mimics. Dual luciferase reporter assay confirmed the binding of circRNA_014511 and mmu-miR-29b-2-5p. Flow cytometry analysis showed the apoptosis rate of bone marrow mesenchymal stem cells overexpressing circRNA_014511 was significantly decreased. In the circRNA_014511 transfection group, after cells were subjected to 6Gy irradiation, G2 phase arrest appeared, the expression of P21 and GADD45A was significantly decreased, and cyclin B1 was significantly increased. Colony formation assay showed the survival fraction of circRNA_014511 overexpression cells after irradiation was significantly higher than control group, and the radiosensitivity was decreased. In conclusion,our findings demonstrated that circRNA_014511 could inhibit the expression of P53 by binding miR-29b-2-5p, and decrease the radiosensitivity of bone marrow mesenchymal stem cells by affecting cell cycle and cell apoptosis.

摘要

造血干细胞移植常用于某些血液系统或骨髓肿瘤患者。全身照射联合化疗是预处理方案的一部分,该方案在造血干细胞移植前最常使用。然而,全身照射预处理会损伤骨髓中的其他正常细胞。因此,探索骨髓间充质干细胞辐射抗性的机制对于细胞移植后恢复造血功能具有重要意义。本研究旨在证明 circRNA_014511 吸附 miR-29b,并探讨 circRNA_014511 对骨髓间充质干细胞放射敏感性的影响。在本研究中,构建了 circRNA_014511 过表达载体,并转染入骨髓间充质干细胞,发现 miR-29b-2-5p 和 P53 降低,这可以通过 miR29b-mimics 逆转。双荧光素酶报告基因实验证实了 circRNA_014511 与 mmu-miR-29b-2-5p 的结合。流式细胞术分析显示,过表达 circRNA_014511 的骨髓间充质干细胞的凋亡率显著降低。在 circRNA_014511 转染组中,细胞经 6Gy 照射后出现 G2 期阻滞,P21 和 GADD45A 的表达显著降低,cyclin B1 显著增加。集落形成实验显示,circRNA_014511 过表达细胞在照射后的存活分数明显高于对照组,放射敏感性降低。总之,我们的研究结果表明,circRNA_014511 通过结合 miR-29b-2-5p 抑制 P53 的表达,并通过影响细胞周期和细胞凋亡降低骨髓间充质干细胞的放射敏感性。

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