程序性细胞死亡因子4增强三氧化二砷对神经母细胞瘤细胞生长及核因子κ B信号通路的抑制作用  

PDCD4 enhances the inhibitory effect of As2O3 on the growth and NF-κB signaling pathway in neuroblastoma cells

作　　者：刘慧娟[1] 李桂玲[1] 雷平冲[2] Liu Huijuan;Li Guiling;Lei Pingchong(Department of Pediatrics, Henan Provincial People′s Hospital, Zhengzhou 450000, China;Department of Hematology, Henan Provincial People′s Hospital, Zhengzhou 450000, China)

机构地区：[1]河南省人民医院儿科,郑州450000 [2]河南省人民医院血液内科,郑州450000

出　　处：《中华肿瘤杂志》2019年第9期675-680,共6页Chinese Journal of Oncology

摘　　要：目的探讨程序性细胞死亡因子4(PDCD4)对三氧化二砷(As2O3)诱导的神经母细胞瘤细胞生长和核因子κB(NF-κB)信号通路的抑制作用。方法在神经母细胞瘤细胞中转染PDCD4过表达载体,以荧光定量PCR和Western blot方法测定过表达效果。将神经母细胞瘤细胞分为对照组、PDCD4组、As2O3组和As2O3+PDCD4组,以四甲基偶氮唑蓝法测定细胞的增殖能力,克隆形成实验测定细胞克隆形成能力,流式细胞术测定细胞凋亡变化,Western blot法测定细胞中NF-κB p65和激活型caspase-3(cleaved caspase-3)蛋白的表达水平。结果PDCD4过表达载体转染可以明显提高神经母细胞瘤细胞中PDCD4的表达水平。对照组、PDCD4组、As2O3组和As2O3+PDCD4组细胞的存活率分别为100%、(72.14±5.20)%、(62.58±3.14)%和(40.87±2.47)%,克隆形成率分别为(91.25±8.36)%、(65.32±7.14)%、(57.23±5.28)%和(37.14±3.64)%,凋亡率分别为(3.57±0.24)%、(28.64±3.20)%、(36.41±4.58)%和(49.65±5.27)%。与对照组比较,PDCD4组、As2O3组和As2O3+PDCD4组细胞的存活率、克隆形成率均明显降低(均P<0.05),凋亡率明显升高(均P<0.05),As2O3+PDCD4组较PDCD4组和As2O3组变化更为显著(均P<0.05)。对照组、PDCD4组、As2O3组和As2O3+PDCD4组cleaved caspase-3蛋白的表达水平分别为0.21±0.03、0.30±0.02、0.43±0.05和0.57±0.06,NF-κB p65蛋白的表达水平分别为0.68±0.04、0.52±0.03、0.43±0.04和0.32±0.02。与对照组比较,PDCD4组、As2O3组和As2O3+PDCD4组细胞中NF-κB p65蛋白的表达水平均明显降低(均P<0.05),cleaved caspase-3蛋白的表达水平均明显升高(P<0.05),As2O3+PDCD4组较PDCD4组和As2O3组变化更为显著(均P<0.05)。结论PDCD4可增强As2O3对神经母细胞瘤细胞生长和NF-κB信号通路的抑制作用。Objective To investigate the inhibitory effect of programmed cell death factor 4 (PDCD4) on arsenic trioxide (As2O3)-induced cell growth and nuclear factor kappa B (NF-κB) signaling pathway in neuroblastoma. Methods The PDCD4 overexpression vector was transfected into neuroblastoma cells and detected by fluorescence quantitative PCR and Western blot. As2O3 was used to treat PDCD4 overexpressing neuroblastoma cells. MTT assay was used to measure the proliferation. Colony formation assay was used to determine the cell clone forming ability. Apoptosis was measured by flow cytometry. Western blot was used to detect the expression of NF-κB p65 and cleaved caspase-3 protein in cells. Results The transfection of PDCD4 overexpression vector significantly increased the expression level of PDCD4 in neuroblastoma cells. The cell survival rates of the control group, PDCD4 group, As2O3 group and As2O3+ PDCD4 group were 100%,(72.14±5.20)%,(62.58±3.14)% and (40.87±2.47)%, respectively. The colony formation rates in these four groups were (91.25±8.36)%,(65.32±7.14)%,(57.23±5.28)% and (37.14±3.64)%, respectively. In addition, the cell apoptotic rates of these four groups were (3.57±0.24)%,(28.64±3.20)%,(36.41±4.58)% and (49.65±5.27)%, respectively. Therefore, overexpression of PDCD4 in the absence or presence of As2O3 inhibited cell proliferation and clone formation ability, while promoted apoptosis. Furthermore, the expression levels of cleaved caspase-3 in the control group, PDCD4 group, As2O3 group and As2O3+ PDCD4 group were 0.21±0.03, 0.30±0.02, 0.43±0.05 and 0.57±0.06, respectively. And the expression levels of NF-κB p65 protein were 0.68±0.04, 0.52±0.03, 0.43±0.04, and 0.32±0.02, respectively. Compared with the control group, the expression levels of NF-κB p65 protein in PDCD4 group, As2O3 group and As2O3+ PDCD4 group were significantly decreased (P<0.05), whereas the expression level of cleaved Caspase-3 protein was significantly increased (P<0.05). The changes in As2O3+ PDCD4 group were more signif

关 键 词：神经母细胞瘤 程序性细胞死亡因子4 核转录因子ΚB 信号通路 

分 类 号：R739.4[医药卫生—肿瘤]