lncRNA SBF2-AS1 affects the proliferation, apoptosis and radiosensitivity of glioma cells by regulating the miR-1287-5p/FSCN1 axis
10.3760/cma.j.cn112152-20200803-00703
- VernacularTitle:SBF2－AS1通过调控miR－1287－5p/FSCN1轴影响胶质瘤细胞的增殖凋亡和放射敏感性
- Author:
Yinghai JIANG
1
;
Lingjie XIA
;
Chaoyue LI
;
Haiqin LI
Author Information
1. 河南省人民医院疼痛科，郑州 450000
- Keywords:
Glioma;
SET binding factor 2 antisense RNA 1;
MiR-1287-5p;
Fascin 1;
Cell proliferation;
Apoptosis;
Radiosensitivity
- From:
Chinese Journal of Oncology
2022;44(8):826-835
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the effect and mechanism of lncRNA SBF2-AS1 on glioma cell proliferation, apoptosis and radiosensitivity.Methods:Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the mRNA expression levels of SBF2-AS1, miR-1287-5p and FSCN1 in normal human brain glial cell HEB and glioma cell lines, including LN18, SW1088, Hs683, and western blot method was used to detect the expression level of FSCN1 protein in cells. Glioma cells including LN18 and SW1088 were taken as the research object. After SBF2-AS1 small interfering RNA, miR-1287-5p mimics, FSCN1 small interfering RNA were transfected into LN18 cells, CCK-8 was used to detect cell proliferation, flow cytometry was used to detect cell apoptosis, clone formation experiment was used to detect cell radiosensitivity, Western blot was used to detect the protein expressions of cyclin D1, cleaved-caspase 3 and phosphorylated histone 2A variant phosphorylated histone (γ-H2AX). Dual luciferase reporter gene experiments verified the regulatory relationship between SBF2-AS1 and miR-1287-5p, as well as miR-1287-5p and FSCN1.Results:Compared with HEB cells, the expression level of SBF2-AS1 and the expression levels of FSCN1 mRNA and protein in glioma cell line LN18, SW1088 and Hs683 were significantly increased ( P<0.05), and the expression level of miR-1287-5p was significantly reduced ( P<0.05). After down-regulating SBF2-AS1, up-regulating miR-1287-5p or down-regulating FSCN1 expression, LN18 and SW1088 cells activity and cyclin D1 protein expression were significantly reduced ( P<0.05), the apoptosis rate and cleared-caspase-3 protein expression were significantly increased ( P<0.05), the survival score was significantly reduced ( P<0.05), and the expression ofγ-H2AX protein was significantly increased ( P<0.05). The results of dual luciferase reporter gene assay showed that the luciferase activity of LN18 and SW1088 cells co-transfected with miR-1287-5p mimics and SBF2-AS1-WT or FSCN1-WT was lower than that of co-transfected miR-NC and SBF2-AS1-WT or FSCN1-WT in LN18 and SW1088 cells ( P<0.001), while the luciferase activity of LN18 and SW1088 cells co-transfected with miR-1287-5p mimics and SBF2-AS1-MUT or FSCN1-MUT was not significantly different from that of miR-NC transfected with SBF2-AS1-MUT or FSCN1-MUT ( P>0.05). The expression level of miR-1287-5p in LN18 and SW1088 cells in si-SBF2-AS1 group was higher than that in si-NC group ( P<0.05), and the expression level of miR-1287-5p in LN18 and SW1088 cells in pcDNA-SBF2-AS1 group was higher than that in si-NC group ( P<0.05), but lower than that of pcDNA-NC group ( P<0.05). The protein expression level of FSCN1 in LN18 and SW1088 cells in the miR-1287-5p group was significantly lower than that in the miR-NC group ( P<0.05), and the protein expression level of FSCN1 in LN18 and SW1088 cells in the anti-miR-1287-5p group was significantly higher than that in the anti-miR-NC group ( P<0.05). When miR-1287-5p and SBF2-AS1 were down-regulated simultaneously or FSCN1 was up-regulated while SBF2-AS1 was down-regulated simultaneously, the proliferation and cyclin D1 protein expression of LN18 and SW1088cells were significantly increased ( P<0.001), the apoptosis rate and cleared-caspase-3 protein expression were significantly reduced ( P<0.001), the survival score was significantly increased ( P<0.001), and the expression of γ-H2AX protein was significantly reduced ( P<0.001). Conclusion:SBF2-AS1 highly expresses in glioma cells, down-regulation of SBF2-AS1 expression can inhibit the proliferation of glioma cells, promote apoptosis, and enhance cell radiosensitivity by regulating the miR-1287-5p/FSCN1 axis.
