Selenoprotein T Promotes Proliferation and G1-to-S Transition in SK-N-SH Cells: Implications in Parkinson's Disease
Selenoprotein T Promotes Proliferation and G1-to-S Transition in SK-N-SH Cells: Implications in Parkinson's Disease
复制标题
硒蛋白 T 促进 SK-N-SH 细胞增殖和 G1 至 S 转变：对帕金森病的影响。
DOI：
10.1093/jn/nxz199
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发表时间：
2019-12-01
影响因子：
4.2
通讯作者：
Zhu, Jian-Hong
中科院分区：
文献类型：
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作者：
Shao, Zi-Qiang;Zhang, Xiong;Zhu, Jian-Hong
背景：硒优先进入大脑，主要用于硒蛋白的表达。硒蛋白T（SELENOT）对帕金森病（PD）小鼠模型中多巴胺能神经元的保护作用目的：我们假设SELENOT在神经细胞中具有增殖作用。为了评估PD中的SELENOT状态，在10-12周龄的镇静雄性C57 BL/6小鼠的神经元中注射6-羟基多巴胺，从9名健康受试者（56%男性，68岁）和11名患有PD的受试者（64%男性，63岁）中分离人外周血单核细胞。多巴胺能神经祖细胞样SK-N-SH细胞与短暂的SELENOT过表达或敲低，保持在存在或不存在的抗氧化剂N-乙酰-L-半胱氨酸和钙通道阻滞剂尼莫地平。结果：经碘化1-甲基-4-苯基吡啶鎓处理的SK-N-SH细胞和PD患者外周血单个核细胞中SELENOT mRNA丰度分别增加了3.5倍和1.6倍（P < 0.05）。同样，在注射6-羟基多巴胺的小鼠的酪氨酸羟化酶阳性多巴胺能神经元中表达了SELENOT。在SK-N-SH细胞中敲低SELENOT抑制（54%; P < 0.05）5-乙炔基-2 '-脱氧尿苷掺入，但诱导（17-47%; P < 0.05）膜联蛋白V阳性细胞、半胱氨酸天冬氨酸蛋白酶-3切割和G1/S细胞周期停滞。SELENOT敲低和过表达分别增加（88-120%; P < 0.05）和减少（37-42%; P < 0.05）叉头盒O3和p27，但减少（51%; P < 0.05）和增加（1.2倍; P < 0.05）细胞周期蛋白依赖性激酶4蛋白丰度。尼莫地平或N-乙酰-L-半胱氨酸治疗（24小时）在稳态水平，这些蛋白质的变化减少。虽然N-乙酰-L-半胱氨酸治疗并没有影响的减少量的钙（13%; P < 0.05）的SELENOT敲低，尼莫地平治疗逆转了减少量的活性氧（33%; P < 0.05）的SELENOT overexpression.Conclusions：这些细胞和小鼠数据链接SELENOT神经增殖，扩大了我们的理解硒保护PD。
Background: Selenium is prioritized to the brain mainly for selenoprotein expression. Selenoprotein T (SELENOT) protects dopaminergic, postmitotic neurons in a mouse model of Parkinson's disease (PD).Objective: We hypothesized a proliferative role of SELENOT in neural cells.Methods: To assess SELENOT status in PD, sedated male C57BL/6 mice at 10-12 wk of age were injected with 6-hydroxydopamine in neurons, and human peripheral blood mononuclear cells were isolated from 9 healthy subjects (56% men, 68-y-old) and 11 subjects with PD (64% men, 63-y-old). Dopaminergic neural progenitor-like SK-N-SH cells with transient SELENOT overexpression or knockdown were maintained in the presence or absence of the antioxidant N-acetyl-L-cysteine and the calcium channel blocker nimodipine. Cell cycle, proliferation, and signaling parameters were determined by immunoblotting, qPCR, and flow cytometry.Results: SELENOT mRNA abundance was increased (P < 0.05) in SK-N-SH cells treated with 1-methyl-4-phenylpyridinium iodide (3.5-fold) and peripheral blood mononuclear cells from PD patients (1.6-fold). Likewise, SELENOT was expressed in tyrosine hydroxylase-positive dopaminergic neurons of 6-hydroxydopamine-injected mice. Knockdown of SELENOT in SK-N-SH cells suppressed (54%; P < 0.05) 5-ethynyl-2'-deoxyuridine incorporation but induced (17-47%; P < 0.05) annexin V-positive cells, CASPASE-3 cleavage, and G1/S cell cycle arrest. SELENOT knockdown and overexpression increased (88-120%; P < 0.05) and reduced (37-42%; P < 0.05) both forkhead box O3 and p27, but reduced (51%; P < 0.05) and increased (1.2-fold; P < 0.05) cyclin-dependent kinase 4 protein abundance, respectively. These protein changes were diminished by nimodipine or N-acetyl-L-cysteine treatment (24 h) at steady-state levels. While the N-acetyl-L-cysteine treatment did not influence the reduction in the amount of calcium (13%; P < 0.05) by SELENOT knockdown, the nimodipine treatment reversed the decreased amount of reactive oxygen species (33%; P < 0.05) by SELENOT overexpression.Conclusions: These cellular and mouse data link SELENOT to neural proliferation, expanding our understanding of selenium protection in PD.