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.2019 May 20;19(2):155-163.
doi: 10.17305/bjbms.2019.3935.

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

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

Yanjie Wang et al. Bosn J Basic Med Sci..

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.

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Figures

FIGURE 1
FIGURE 1
The regulation of p53 expression by the combination of circRNA_014511 and mmu-miR-29b-2-5p in BMMSCs. (A) and (B), quantitative fluorescence PCR detection of the expressions of circRNA_014511 and mmu-miR-29b-2-5p in the cells of each group. (C) and (D) results of dual luciferase reporter assay. (E) and (F) Western blot assessment of p53 expression.
FIGURE 2
FIGURE 2
The effect of circRNA_014511 on apoptosis of BMMSCs. (A) Western blot assessment of the expressions of apoptosis-related proteins p53, Bcl-2, and Mcl-1. (B) Flow cytometry detection of apoptosis in cells of each group.
FIGURE 3
FIGURE 3
The effect of circRNA_014511 on cell cycle of BMMSCs. (A) Flow cytometry assessment of cell cycle in cells of each group.(B) Western blot assessment of the expressions of cell cycle-related proteins P21, GADD45A, and Cyclin B1.
FIGURE 4
FIGURE 4
Effect of circRNA_014511 on the radiosensitivity of BMMSCs.
See this image and copyright information in PMC

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