Ishido Masami
Endocrine Disruptors Research Projects, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba 305-8506, Japan.
Regul Pept. 2005 Jan 15;124(1-3):45-51. doi: 10.1016/j.regpep.2004.06.027.
The addition of tumor necrosis factor (TNF)-alpha into the cultured porcine kidney LLC-PK1 cells caused apoptosis concomitantly with caspase-3 activation and the inductions of an endogenous Bcl-2 protein. An SDS-polyacrylamide electrophoretic analysis revealed that a 37-kDa protein in a nuclear fraction was increased during TNF-alpha-induced apoptosis. Partial amino acid sequence of the protein was A-L-T-G-H-L-E-E-V, perfectly matching that of annexin I. Immunocytochemistry revealed that annexin I migrated to the nucleus and/or peri-nucleus region upon exposure to TNF-alpha. Overexpression of Bcl-2 proteins inhibited the nuclear localization of annexin I during TNF-alpha-induced apoptosis. Antisense oligodeoxynucleotides complementary to annexin I-inhibited TUNEL (terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labelling) staining in TNF-alpha-treated cells, suggesting that annexin I expression is a possible prerequisite for the induction of apoptosis by the cytokine. Thus, it is first time to show that annexin I is regulated by an anti-apoptotic Bcl-2 protein in TNF-alpha-induced renal apoptotic events.
在培养的猪肾LLC-PK1细胞中添加肿瘤坏死因子（TNF）-α会导致细胞凋亡，同时伴有半胱天冬酶-3的激活和内源性Bcl-2蛋白的诱导。十二烷基硫酸钠-聚丙烯酰胺电泳分析显示，在TNF-α诱导的细胞凋亡过程中，核组分中的一种37 kDa蛋白增加。该蛋白的部分氨基酸序列为A-L-T-G-H-L-E-E-V，与膜联蛋白I完全匹配。免疫细胞化学显示，在暴露于TNF-α后，膜联蛋白I迁移至细胞核和/或核周区域。在TNF-α诱导的细胞凋亡过程中，Bcl-2蛋白的过表达抑制了膜联蛋白I的核定位。与膜联蛋白I互补的反义寡脱氧核苷酸抑制了TNF-α处理细胞中的TUNEL（末端脱氧核苷酸转移酶（TdT）介导的dUTP缺口末端标记）染色，表明膜联蛋白I的表达可能是该细胞因子诱导细胞凋亡的一个前提条件。因此，首次表明在TNF-α诱导的肾细胞凋亡事件中，膜联蛋白I受抗凋亡Bcl-2蛋白的调节。
