TISSUE TRANSGLUTAMINASE AND APOPTOSIS - SENSE AND ANTISENSE TRANSFECTION STUDIES WITH HUMAN NEUROBLASTOMA-CELLS
TISSUE TRANSGLUTAMINASE AND APOPTOSIS - SENSE AND ANTISENSE TRANSFECTION STUDIES WITH HUMAN NEUROBLASTOMA-CELLS
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DOI：
10.1128/mcb.14.10.6584
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发表时间：
1994-10-01
影响因子：
5.3
通讯作者：
PIACENTINI, M
中科院分区：
文献类型：
--
作者：
MELINO, G;ANNICCHIARICOPETRUZZELLI, M;PIACENTINI, M
在这份报告中，我们表明，在人神经母细胞瘤细胞系SK-N-BE（2）中组织转氨酶（tTG）的过度表达使这些神经嵴来源的细胞对凋亡死亡高度敏感。在组成型启动子的控制下，用全长tTG cDNA转染的细胞显示出增殖能力的急剧降低，同时细胞死亡率大幅增加。垂死的tTG转染细胞表现出细胞凋亡的细胞质和细胞核的变化特征。tTG转染的细胞表达高Bcl-2蛋白水平以及沿着神经元途径分化的细胞的表型神经细胞粘附分子标记物（NCAM和神经丝）。与这些研究结果一致，用含有反义方向的人tTG cDNA片段的表达载体转染神经母细胞瘤细胞导致自发和视黄酸（RA）诱导的细胞凋亡显著减少。我们还提供了证据表明：（i）这些神经外胚层细胞的凋亡程序受到RA的严格调控;（ii）在人神经母细胞瘤SK-N-BE（2）细胞中，细胞凋亡导致的细胞死亡优先发生在基质粘附表型中。这是第一次，我们在这里报告tTG在细胞凋亡的表型成熟的直接影响。这些结果表明，细胞内蛋白质的TG依赖性不可逆交联是诱导细胞凋亡导致结构变化的重要生化事件。
In this report, we show that the overexpression of tissue transglutaminase (tTG) in the human neuroblastoma cell line SK-N-BE(2) renders these neural crest-derived cells highly susceptible to death by apoptosis. Cells transfected with a full-length tTG cDNA, under the control of a constitutive promoter, show a drastic reduction in proliferative capacity paralleled by a large increase in cell death rate. The dying tTG-transfected cells exhibit both cytoplasmic and nuclear changes characteristic of cells undergoing apoptosis. The tTG-transfected cells express high Bcl-2 protein levels as well as phenotypic neural cell adhesion molecule markers (NCAM and neurofilaments) of cells differentiating along the neuronal pathway. In keeping with these findings, transfection of neuroblastoma cells with an expression vector containing segments of the human tTG cDNA in antisense orientation resulted in a pronounced decrease of both spontaneous and retinoic acid (RA)-induced apoptosis. We also present evidence that (i) the apoptotic program of these neuroectodermal cells is strictly regulated by RA and (ii) cell death by apoptosis in the human neuroblastoma SK-N-BE(2) cells preferentially occurs in the substrate-adherent phenotype. For the first time, we report here a direct effect of tTG in the phenotypic maturation toward apoptosis. These results indicate that the tTG-dependent irreversible cross-linking of intracellular protein represents an important biochemical event in the induction of the structural changes featuring cells dying by apoptosis.