...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of cellular biochemistry. >Overexpression of regucalcin suppresses apoptotic cell death in cloned normal rat kidney proximal tubular epithelial NRK52E cells: Change in apoptosis-related gene expression.
【24h】

Overexpression of regucalcin suppresses apoptotic cell death in cloned normal rat kidney proximal tubular epithelial NRK52E cells: Change in apoptosis-related gene expression.

机译：regucalcin的过表达抑制克隆的正常大鼠肾近端肾小管上皮NRK52E细胞中凋亡细胞死亡：凋亡相关基因表达的变化。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The effect of regucalcin, a regulatory protein in intracellular signaling pathway, on cell death and apoptosis was investigated using the cloned normal rat kidney proximal tubular epithelial NRK52E cells overexpressing regucalcin. NRK52E cells (wild type) and stable regucalcin (RC)/pCXN2 transfectants were cultured for 72 h in a medium containing 5% bovine serum (BS) to obtain subconfluent monolayers. After culture for 72 h, cells were further cultured for 24-72 h in a medium without BS containing either vehicle, tumor necrosis factor-alpha (TNF-alpha; 0.1 or 1.0 ng/ml of medium), lipopolysaccharide (LPS; 0.1 or 1.0 microg/ml), Bay K 8644 (10(-9)-10(-7) M), or thapsigargin (10(-9)-10(-7) M). The number of wild-type cells was significantly decreased by culture for 42-72 h in the presence of TNF-alpha (0.1 or 1.0 ng/ml), LPS (0.1 or 1.0 microg/ml), Bay K 8644 (10(-7)-10(-5) M), or thapsigargin (10(-8) or 10(-7) M). The effect of TNF-alpha (0.1 or 1.0 ng/ml), LPS (0.1 or 1.0 microg/ml), Bay K 8644 (10(-7)-10(-6) M), or thapsigargin (10(-7) M) in decreasing the number of wild-type cells cultured for 24-72 h was significantly prevented in transfectants overexpressing regucalcin. Agarose gel electrophoresis showed the presence of low-molecular-weight deoxyribonucleic acid (DNA) fragments of adherent wild-type cells cultured with LPS (1.0 microg/ml), Bay K 8644 (10(-7) M), or thapsigargin (10(-8) M) for 24 h, and this DNA fragmentation was significantly suppressed in transfectants. DNA fragmentation in adherent cells was not seen by culture with TNF-alpha (1.0 ng/ml). TNF-alpha-induced decrease in the number of wild-type cells was significantly prevented by culture with caspase-3 inhibitor (10(-8) M), while LPS- or Bay K 8644-induced decrease in cell number was significantly prevented by caspase-3 inhibitor or N omega-nitro-L-arginine methylester (NAME) (10(-5) M), an inhibitor of nitric oxide (NO) synthase. Thapsigargin-induced decrease in cell number was not prevented in the presence of two inhibitors. Bcl-2 and Akt-1 mRNA levels were significantly increased in transfectants cultured for 24 h as compared with those of wild-type cells, while Apaf-1, caspase-3, or glyceroaldehyde-3-phosphate dehydrogenase (G3PDH) mRNA expressions were not significantly changed in transfectants. Culture with TNF-alpha (1.0 ng/ml), LPS (1.0 microg/ml), Bay K 8644 (l0(-7) M), or thapsigargin (10(-8) M) caused a significant increase in caspase-3 mRNA levels in wild-type cells. LPS (1.0 microg/ml) significantly decreased Bcl-2 mRNA expression in the cells. Their effects on the gene expression of apoptosis-related proteins were not significantly changed in transfectants. This study demonstrates that overexpression of regucalcin has a suppressive effect on cell death and apoptosis induced by various factors which their action are mediated through many intracellular signaling pathways, and that it modulates the gene expression of apoptosis-related proteins. J. Cell. Biochem. (c) 2005 Wiley-Liss, Inc.
机译：使用克隆的正常大鼠肾脏近端肾小管上皮NRK52E细胞过表达的regucalcin，研究了regucalcin（一种细胞内信号传导途径中的调节蛋白）对细胞死亡和凋亡的影响。将NRK52E细胞（野生型）和稳定的regucalcin（RC）/ pCXN2转染子在含有5％牛血清（BS）的培养基中培养72小时，以获得亚汇合单层。培养72小时后，将细胞在不含BS的培养基中进一步培养24-72小时，所述培养基不含运载体，肿瘤坏死因子-α（TNF-α; 0.1或1.0ng / ml的培养基），脂多糖（LPS; 0.1或1.0）。 1.0微克/毫升），Bay K 8644（10（-9）-10（-7）M）或毒胡萝卜素（10（-9）-10（-7）M）。在TNF-α（0.1或1.0 ng / ml），LPS（0.1或1.0 microg / ml），Bay K 8644（10（- 7）-10（-5）M）或毒胡萝卜素（10（-8）或10（-7）M）。 TNF-alpha（0.1或1.0 ng / ml），LPS（0.1或1.0 microg / ml），Bay K 8644（10（-7）-10（-6）M）或thapsigargin（10（-7 ）M）在过量表达regucalcin的转染子中可显着防止减少培养24-72 h的野生型细胞数量。琼脂糖凝胶电泳显示存在低分子量的脱氧核糖核酸（DNA）片段，这些片段是用LPS（1.0 microg / ml），Bay K 8644（10（-7）M）或thapsigargin（10 （-8）M）24 h，并且该DNA片段在转染子中被显着抑制。用TNF-α（1.0 ng / ml）培养未见贴壁细胞的DNA断裂。用caspase-3抑制剂（10（-8）M）培养可显着防止TNF-α诱导的野生型细胞数量减少，而LPS或Bay K 8644诱导的细胞数量减少则可通过以下方式显着防止caspase-3抑制剂或一氧化氮（NO）合酶抑制剂Nω-硝基-L-精氨酸甲酯（NAME）（10（-5）M）。在两种抑制剂的存在下，毒胡萝卜素诱导的细胞数量减少并未得到阻止。与野生型细胞相比，培养24 h的转染子中Bcl-2和Akt-1 mRNA水平显着增加，而Apaf-1，caspase-3或甘油醛-3-磷酸脱氢酶（G3PDH）mRNA表达则明显升高。转染子没有明显改变。与TNF-alpha（1.0 ng / ml），LPS（1.0 microg / ml），Bay K 8644（10（-7）M）或thapsigargin（10（-8）M）的培养引起caspase-3的显着增加。野生型细胞中的mRNA水平。 LPS（1.0微克/毫升）显着降低了细胞中Bcl-2 mRNA的表达。在转染子中它们对凋亡相关蛋白基因表达的影响没有明显改变。这项研究表明，regucalcin的过表达对多种因素诱导的细胞死亡和凋亡具有抑制作用，而这些因素的作用是通过许多细胞内信号途径介导的，并且它调节凋亡相关蛋白的基因表达。 J.细胞。生化。 （c）2005 Wiley-Liss，Inc.

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号