Effect and mechanism of the role of Sal-miR-58 in mediating radiosensitivity of glioma U251 cells via HAX1
10.3760/cma.j.cn113030-20231007-00117
- VernacularTitle:Sal-miR-58通过HAX1介导胶质瘤细胞放射敏感性的作用及机制研究
- Author:
Guohui WANG
1
;
Hongyao GE
;
Zhenyu DU
;
Gaoshan YANG
Author Information
1. 天津市第一中心医院放疗科，天津 300000
- Keywords:
MicroRNAs, salvia miltiorrhiza-derived;
HAX1 gene;
Radiosensitivity;
Glioma cells
- From:
Chinese Journal of Radiation Oncology
2024;33(8):746-752
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of salvia miltiorrhiza-derived Sal-miR-58 on the radiosensitivity of glioma U251 cells and its possible mechanism. Methods:Glioma U251 cells were treated with different concentrations of miR-Ctl or Sal-miR-58 mimic, and subsequently treated with radiation to establish the radiotherapy model in vitro. The effect of Sal-miR-58 upon U251 cell viability was assessed by MTT assay. The effect of Sal-miR-58 on apoptosis of glioma U251 cells was evaluated by Hoechst33342/ propidium iodide (PI) staining. The changes of reactive oxygen species (ROS) content in U251 cells were detected by 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe. The effect of Sal-miR-58 combined irradiation on the radiosensitivity of U251 cells was detected by clone formation assay. The expression levels of HCLS1-related protein X-1 (HAX1), apoptosis marker proteins B cell lymphoma 2 (Bcl-2), cleaved-cysteine-containing aspartate-specific protease (Cleaved-Caspase) 9 and Cleaved-Caspase 3 were detected by Western blot. Multi-group comparison was conducted by one-way ANOVA. Two-group comparison was performed by independent sample t-test. Results:Sal-miR-58 could exacerbate the inhibition of U251 cell proliferation after irradiation ( P<0.05). Sal-miR-58 could promote the apoptosis of U251 cells and increase the production of ROS in U251 cells after radiation ( P<0.05). Clone formation assay showed that Sal-miR-58 increased the radiosensitivity of U251 cells, with a radiosensitization ratio of 1.43. Western blot showed that Sal-miR-58 inhibited the expression of HAX1 in U251 cells. Sal-miR-58 could inhibit the expression of Bcl-2 and increase the activation of Caspase 3 and Caspase 9 by reducing the expression of HAX1. Conclusions:Sal-miR-58 enhances the radiosensitivity of U251 cells. The possible mechanism is that Sal-miR-58 inhibits the expression of HAX1 induced by radiation and accelerates the apoptosis process of tumor cells.
