摘要目的:探讨长链非编码RNA(lncRNA)Sox2OT过表达对Aβ1-42诱导PC12细胞损伤及PI3K/Akt通路的影响。方法:选取大鼠嗜铬神经瘤细胞株(PC12)，利用Aβ1-42对PC12细胞进行处理，建立AD的细胞模型。将Aβ1-42诱导前的PC12细胞设为空白组；将Aβ1-42诱导后的PC12细胞分为control组、Sox2OT过表达(p-Sox2OT)组、p-Sox2OT空载体(p-NC)组、抑制Sox2OT表达(si-Sox2OT)组和si-Sox2OT空载体(si-NC)组。使用噻唑蓝(MTT)方法对细胞的增殖活性进行检测；采用流式细胞术对转染后的细胞周期和凋亡率进行检测；采用Western blot检测PI3K/Akt通路蛋白的表达。结果:MTT结果显示，与空白组(99.67±10.50 )相比，control组(29.33±5.51 )的细胞增殖率显著降低( t＝10.27， P<0.05)。RT-qPCR结果显示，与control组(0.52±0.06)相比，p-Sox2OT组(2.19±0.16)中的Sox2OT的mRNA表达水平显著升高( t＝16.93， P<0.05)，si-Sox2OT组(0.22±0.02)中的Sox2OT的mRNA表达水平显著下降( t＝15.28， P<0.05)；与p-NC组(0.53±0.12)相比，p-Sox2OT组(2.19±0.16)中的Sox2OT的mRNA表达水平显著升高( t＝16.25， P<0.05)；与si-NC组(0.51±0.09)相比，si-Sox2OT组(0.22±0.02)中的Sox2OT的mRNA表达水平显著下降( t＝16.93， P<0.05)；control组与p-NC组以及si-NC组间的差异无统计学意义( P>0.05)。此外，p-Sox2OT组细胞的增殖能力(145.00±5.12)显著高于si-Sox2OT组(23.33±4.93)、control组(55.00±5.00)、si-NC组(57.33±8.51)以及p-NC组(56.00±5.57)( t＝29.65，21.78，27.55，21.35，均 P<0.05)。Control组与p-NC组以及si-NC组间细胞增殖率的差异不具有统计学意义( P>0.05)。细胞周期检测实验显示，p-Sox2OT组G1期的细胞数量显著低于control组和p-NC( t＝9.80，8.57；均 P<0.05)，而p-Sox2OT组G2期的细胞数量与control组和p-NC组相比却显著升高( t＝11.02，10.25；均 P<0.05)；si-Sox2OT组G1期的细胞数量显著高于control组和si-NC组( t＝8.22，3.11，均 P<0.05)，而G2期的细胞数量与control组和si-NC组相比却显著下降( t＝6.32，5.33；均 P<0.05)；control组与p-NC组以及si-NC组间的细胞周期差异无统计学意义(均 P>0.05)。在S期中，只有p-Sox2OT组与control组之间的差异有统计学意义( t＝1.84， P<0.05)。p-Sox2OT组细胞凋亡率[(3.66±0.26)%]低于si-Sox2OT组[(14.25±0.80)%]、control组[(8.46±0.44)%]、si-NC组[(8.78±0.44)%]以及p-NC组[(8.40±0.21)%]( t＝21.81，16.27，20.32，21.35，均 P<0.05)。Control组与p-NC组以及si-NC组间的细胞凋亡率差异不具有统计学意义( P>0.05)。Western blot结果显示，p-Sox2OT组中p-PI3K和p-Akt蛋白的表达明显高于p-NC( P<0.05)；与si-NC组相比，si-Sox2OT组PC12细胞中p-PI3K和p-Akt蛋白的表达显著下降( P<0.05)。 结论:lncRNA Sox2OT可以通过调控PI3K/Akt通路促进Aβ1-42诱导的PC12细胞的增殖，抑制细胞的凋亡。

abstractsObjective: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.