Protective effect and mechanism of lncRNA Sox2OT overexpression on PC12 cells injury induced by Aβ1-42
10.3760/cma.j.cn371468-20200305-01092
- VernacularTitle:长链非编码RNA Sox2OT过表达对Aβ1-42诱导PC12细胞损伤的保护作用及机制
- Author:
Peng CHEN
1
;
Long LI
;
Chunhua LIU
;
Zhaoqiang ZHANG
;
Xianchang SUN
Author Information
1. 山东第一医科大学（山东省医学科学院）基础医学院生理学教研室，泰安 271016
- From:
Chinese Journal of Behavioral Medicine and Brain Science
2020;29(9):785-791
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the mechanism of lncRNA Sox2OT in patients with Alzheimer's disease (AD) induced by A type of β peptide (Aβ1-42).Methods:Rat pheochromocytoma cells (PC12 cells) were selected and treated by Aβ1-42 to establish PC12 cell model.PC12 cells were set as blank group before induction to verify the successful construction of the cell model.The induced PC12 cells were divided into control group, Sox2OT overexpression (p-Sox2OT) group, p-Sox2OT empty vector (p-NC) group, inhibited Sox2OT expression (si-Sox2OT) group and si-Sox2OT empty vector (si-NC) group.The proliferation activity of thiazole blue (MTT) was detected.Flow cytometry was used to detect the cell cycle and apoptosis rate after transfection.Results:MTT results showed that compared with the blank group (99.67±10.50), the cell proliferation rate of the control group (29.33±5.51) was significantly reduced ( t=10.27, P<0.05). RT-qPCR results showed that compared with the control group (0.52±0.06), the Sox2OT mRNA expression level in the p-Sox2OT group (2.19±0.16) was significantly increased ( t=16.93, P<0.05). The mRNA expression level of Sox2OT in the si-Sox2OT group (0.22±0.02) decreased significantly ( t=15.28, P<0.05). Compared with the p-NC group (0.53±0.12), The mRNA expression level of Sox2OT in the p-Sox2OT group (2.19±0.16) was significantly increased ( t=16.25, P<0.05). Compared with the si-NC group (0.51±0.09), the mRNA expression level of Sox2OT in the si-Sox2OT group (0.22±0.02) was significantly decreased ( t=16.93, P<0.05). The difference between the control group, the p-NC group and the si-NC group was not statistically significant ( P>0.05). In addition, the cell proliferation ability of the p-Sox2OT group (145.00±5.12) was significantly higher than that of the si-Sox2OT group (23.33±4.93), control group (55.00±5.00), si-NC group (57.33±8.51) and p-NC group (56.00±5.57) ( t=29.65, 21.78, 27.55, 21.35, all P<0.05). The difference in cell proliferation rate between Control group, p-NC group and si-NC group was not statistically significant ( P>0.05). Cell cycle detection experiments showed that the number of cells in the G1 phase of the p-Sox2OT group was significantly lower than that of the control group and p-NC ( t=9.80, 8.57; both P<0.05), while the number of cells in the G2 phase of the p-Sox2OT group was significantly higher than that of the control group and the p-NC group ( t=11.02, 10.25; both P<0.05). The number of cells in the G1 phase of the si-Sox2OT group was significantly higher than that of the control group and the si-NC group ( t=8.22, 3.11, both P<0.05), while the number of cells in the G2 phase of the si-Sox2OT group decreased significantly, compared with the control group and the si-NC group ( t=6.32, 5.33; all P<0.05). There was no statistically significant difference in cell cycle between the control group, the p-NC group and the si-NC group (both P>0.05). In the S phase, the difference between the p-Sox2OT group and the control group was statistically significant ( t=1.84, P<0.05). The number of cells in the G2 phase of the p-Sox2OT group (19.00±1.00) was significantly higher than that of the si-Sox2OT group (3.33±1.53), the control group (10.00±1.00), si-NC group (8.55±0.73) and p-NC group (7.67±1.53) ( t=14.85, 11.02, 10.23, 10.74, all P<0.05). The apoptosis rate of p-Sox2OT group ((3.66±0.26)%) was lower than that of si-Sox2OT group ((14.25±0.80)%), control group ((8.46±0.44)%), si-NC group ((8.78±0.44)%) and p-NC group ((8.40± 0.21)%) ( t=21.81, 16.27, 20.32, 21.35, all P<0.05). For the apoptosis rate, there was no statistically significant difference between control group, p-NC group and si-NC group( P>0.05). In addition, the expression levels of p-PI3K and p-Akt in the p-Sox2OT group were significantly higher than those in the p-NC group ( P<0.05). Compared with the si-NC group, the expression of p-PI3K and p-Akt in PC12 cells in the si-Sox2OT group was significantly decreased ( P<0.05). Conclusion:lncRNA Sox2OT can promote the proliferation of PC12 cells induced by Aβ1-42 and inhibit apoptosis by regulating the PI3K/Akt pathway.
