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A study about the impaired mitochondrial membrane potential and nuclear body formation, and their relationship with the induction of cell senescence and cell death

线粒体膜电位和核体形成受损及其与诱导细胞衰老和细胞死亡关系的研究

基本信息

批准号：
17590159
负责人：
NISHIO Koji
金额：
$ 2.3万
依托单位：
NAGOYA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

项目摘要

With fluorescently labeled S1-1 isoform specific antibodies and PML isoform specific antibodies, we examined the nuclear bodies of cultured human normal cells (TIG3S), HeLa cells, various normal rat tissues, and various human tumor tissues with the corresponding normal tissues. In young TIG3S cells, S1-1 nuclear bodies were rarely observed, but PML2, 4, 5, 6 nuclear bodies were observed significantly. Furthermore, co-localization of S1-1 and PML6 nuclear bodies was confirmed. In HeLa cells, S1-1 nuclear bodies were hardly observed. In addition, the remarkable cytoplasmic localization of S1-1 and of PML protein, significant nuclear bodies of PML2 and PML6, and rare nuclear bodies of PML4 and PML5 were observed. We confirmed the rare S1-1 nuclear bodies in dividing young TIG3S and HeLa cells. In addition, the S1-1p110 protein levels of young TIG3S and HeLa cells were very low, on the contrary, the S1-1p130 protein was at very high levels. Furthermore, the S1-1p130 level of HeLa cells was 3-fold higher than TIG3S cells. PML and S1-1(RBM10) have many phosphorylation sites by CK2 and PKC, and HeLa cells have significantly higher activity of CK2 than normal cells. Thereby, the PML isoforms which are phosphorylated (S517 of PML) by CK2, degrade in the proteasomes. However, S1-1p130 (RBM10 variant-1) was very stable even in HeLa cells. HeLa and TIG3S cells, which administered with PKC or CK2 inhibitor, significantly developed the S1-1 nuclear bodies. This suggests that the assembly of S1-1 nuclear bodies was suppressed through their phosphorylation by CK2 in the actively dividing cells. In the presence of PKC or CK2 inhibitor, but not CDK2 inhibitor, cell division of HeLa or TIG3S cells was strongly impaired, and the drugs promoted cell death. Our results suggest that S1-1 and PML nuclear bodies regulate the cell growth and promote the cell death pathway

使用荧光标记的S1-1亚型特异性抗体和PML亚型特异性抗体，我们检查了培养的人类正常细胞（TIG3S）、HeLa细胞、各种正常大鼠组织和各种人类肿瘤组织的核体以及相应的正常组织。在年轻的TIG3S细胞中，很少观察到S1-1核体，但明显观察到PML2、4、5、6核体。此外，还证实了 S1-1 和 PML6 核体的共定位。在HeLa细胞中，几乎观察不到S1-1核体。此外，还观察到S1-1和PML蛋白显着的细胞质定位，PML2和PML6显着的核体，以及PML4和PML5的罕见核体。我们证实了分裂中的年轻 TIG3S 和 HeLa 细胞中存在罕见的 S1-1 核体。此外，年轻的TIG3S和HeLa细胞的S1-1p110蛋白水平非常低，相反，S1-1p130蛋白水平非常高。此外，HeLa 细胞的 S1-1p130 水平比 TIG3S 细胞高 3 倍。 PML和S1-1(RBM10)有许多被CK2和PKC磷酸化的位点，HeLa细胞的CK2活性明显高于正常细胞。因此，被 CK2 磷酸化的 PML 同工型（PML 的 S517）在蛋白酶体中降解。然而，S1-1p130（RBM10 变体-1）即使在 HeLa 细胞中也非常稳定。给予 PKC 或 CK2 抑制剂的 HeLa 和 TIG3S 细胞显着发育出 S1-1 核体。这表明，在活跃分裂的细胞中，S1-1 核体的组装通过 CK2 的磷酸化而受到抑制。在存在 PKC 或 CK2 抑制剂但不存在 CDK2 抑制剂的情况下，HeLa 或 TIG3S 细胞的细胞分裂受到严重损害，并且药物促进细胞死亡。我们的结果表明 S1-1 和 PML 核体调节细胞生长并促进细胞死亡途径