利培酮通过影响Wnt/β-catenin信号通路诱导人成骨细胞系hFob1.19凋亡

Risperidone induces apoptosis of human osteoblast cell line hFob1.19 through Wnt/β-catenin signaling pathway

目的探讨利培酮对人成骨细胞系hFob1.19凋亡的影响,并进一步分析其基于Wnt/β-catenin信号通路的分子机制。方法将hFob1.19细胞分为对照组、利培酮干预组(0、5、20、40、60、80、100和120μmol/L)。CCK-8法检测细胞活力;流式细胞测量术检测细胞凋亡率;RT-qPCR及Western blot检测细胞BCL-2、MCL-1、BAX以及β-catenin基因及蛋白表达水平。结果1)与对照组相比,利培酮组细胞活力呈剂量和时间依赖性下降(P<0.05),而细胞凋亡率呈剂量依赖性增加(P<0.05)。2)与对照组相比,利培酮组促凋亡基因BAX水平升高,而抗凋亡基因BCL-2、MCL-1水平降低,同时β-catenin的基因表达降低(P<0.01)。3)与对照组相比,利培酮组促凋亡蛋白BAX、cleaved caspase-3水平升高,而抗凋亡蛋白BCL-2、MCL-1水平降低,同时β-catenin蛋白表达降低(P<0.01)。4)与对照组相比,利培酮组的β-catenin蛋白在细胞核和细胞质表达均降低(P<0.05)。结论利培酮可以诱导hFob1.19细胞凋亡,其机制可能与利培酮抑制β-catenin表达水平以及抑制β-catenin向核内转移从而导致抗凋亡蛋白质和促凋亡蛋白质平衡被打破相关。

Objective To investigate the effect of risperidone on the apoptosis of human osteoblast cell line hFob1.19 and analyze potential molecular mechanism based on Wnt/β-catenin signaling pathway.MethodshFob1.19 cells were divided into control group and risperidone intervention group(0,5,20,40,60,80,100 and 120μmol/L).Cell viability was detected by CCK-8 assay.The apoptosis rate was detected by flow cytometry.RT-qPCR and Western blot was used to detect the mRNA and protein expression of BCL-2,MCL-1,BAX,andβ-catenin.Results 1)Compared with the control group,the cell viability of the risperidone group decreased in a dose-and time-dependent manner(P<0.05),while the apoptosis rate increased in a dose-dependent manner(P<0.05).2)Compared with the control group,the expression of pro-apoptotic gene BAX in risperidone group increased,while the expression of anti-apoptotic genes BCL-2 and MCL-1 decreased,and the expression ofβ-catenin decreased(P<0.01).3)Compared with the control group,the levels of pro-apoptotic proteins BAX and cleaved caspase-3 were increased,the levels of anti-apoptotic proteins BCL-2 and MCL-1 were decreased,and the expression ofβ-catenin protein was decreased in risperidone group(P<0.01).4)Compared with the control group,the expression ofβ-catenin protein in the nucleus and cytoplasm of risperidone group decreased(P<0.05).Conclusions Risperidone induces apoptosis of hFob1.19 cells,and the mechanism may be related to the inhibition ofβ-catenin expression and nuclear translocation ofβ-catenin by risperidone,which leads to the disruption of the balance between anti-apoptotic and pro-apoptotic proteins.