CircRNA-011235 Counteracts The Deleterious Effect Of Irradiation Treatment On Bone Mesenchymal Stem Cells By Regulating The MiR-741-3p/CDK6 Pathway

Wen, Xianhui; Xie, Hebin; Gui, Rong; Nie, Xinmin; Shan, Dongyong; Huang, Rong; Deng, Hongyu; Zhang, Junhua

doi:10.22074/cellj.2022.7697

CircRNA-011235 Counteracts The Deleterious Effect Of Irradiation Treatment On Bone Mesenchymal Stem Cells By Regulating The MiR-741-3p/CDK6 Pathway

Document Type : Original Article

Authors

¹ Department of Blood Transfusion, The Third Xiangya Hospital of Central South University, Changsha, China

² Research and Teaching Department, Changsha Central Hospital, Changsha, Hunan, China

³ The Third Xiangya Hospital of Central South University, Changsha, China

⁴ Hunan Cancer Hospital and The Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha, China

10.22074/cellj.2022.7697

Abstract

Objective: The present work was aimed at uncovering the effect of circRNA-011235 (circ-011235) on irradiation-induced
bone mesenchymal stem cells (BMSCs) injury and its regulatory mechanism, with a view to establish a scientific basis for its possible medical applications.
Materials and Methods: In this experimental study, after irradiation with different doses (0, 2, 4, 6 GY), the relative expression levels of circ-011235, miR-741-3p, and cyclin-dependent kinases 6 (CDK6) were detected in the BMSCs, using the real time-quantitative polymerase chain reaction (RT-qPCR). The overexpression effects of circ-011235 and CDK6 on the cell proliferation in irradiation-treated BMSCs were measured by the Cell Counting Kit-8 (CCK8) assay. And also, their effects on the cell cycle were evaluated by flow cytometry. RT-qPCR and immunoblotting were performed to detect the effects of pcDNA-circ-011235 and pcDNA-CDK6 on the expression of cyclin D1 and cyclindependent kinases 4 (CDK4) at the gene and protein levels, respectively.
Results: Irradiation treatment elevated the expression of circ-011235 and CDK6, but reduced miR-741-3p expression in the BMSCs with a dose-dependent effect. The proliferation of BMSCs was significantly inhibited in the irradiation treatment group, while the overexpression of circ-011235 and CDK6 effectively attenuated this inhibition. Also, overexpression of circ-011235 and CDK6 elevated the expression of cyclin D1 in irradiation-treated BMSCs, but had no
significant effect on the CDK4 expression.
Conclusion: Our results demonstrated that circ-011235 up-regulated the expression of cyclin D1 via miR-741-3p/CDK6 signal pathway, thereby promoting cell cycle progression and proliferation of irradiation-treated BMSCs. This finding suggested circ-011235/ miR-741-3p/CDK6 pathway exerted a protective role in the response to irradiation and will be a potential new target for future research on the mechanism involved in the resistance of BMSCs to radiation.

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