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首页> 外文期刊>Journal of Cellular Physiology >Intracellular apoptosis-inducing factor is induced by a vacuolar type H+-ATPase inhibitor in B lineage cells.
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Intracellular apoptosis-inducing factor is induced by a vacuolar type H+-ATPase inhibitor in B lineage cells.

机译：细胞内凋亡诱导因素是诱发一种空泡的H + atp酶抑制剂在B谱系细胞。

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摘要

We previously reported that concanamycin A, a specific inhibitor of vacuolar type H+-ATPases, induces DNA fragmentation in B cell hybridoma HS-72 cells. In the present study, we found that the cytosol from concanamycin A-treated HS-72 cells had a cytotoxic effect on intact cells in a cell viability assay. While activin A also induced apoptosis in HS-72 cells, the cytosol from activin A-treated HS-72 cells had no effect on cell viability. We purified the cytosol from concanamycin A-treated HS-72 cells by a four-step procedure: ultracentrifugation; HiTrap heparin column chromatography; HiTrap Q column chromatography; and reverse-phase high performance liquid chromatography on a C18 hydrophobic support. The biologically active fraction, which was used as partially purified cytosol, gave a specific band of protein with a molecular mass of 33 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The mechanism of cell death was examined by observing changes in nuclear morphology, an increase in the proportion of fragmented DNA, and the typical ladder pattern of degraded chromosomal DNA, indicating the induction of apoptosis in cells cultured with the partially purified cytosol. The overexpression of human Bcl-2 suppressed apoptosis, indicating that the cytosol from concanamycin A-treated HS-72 cells induces apoptosis by a Bcl-2-inhibiting mechanism. These findings suggest that concanamycin A, a vacuolar type H+-ATPase inhibitor, produces intracellular apoptosis-inducing factor in B cell hybridoma.
机译：我们以前报道,concanamycin,具体的空泡的类型H + atp酶抑制剂,诱导B细胞杂种细胞的DNA碎片hs - 72细胞。的胞质concanamycin经处理的hs - 72细胞对完整细胞有细胞毒性的影响细胞的可行性分析。诱导细胞凋亡在hs - 72细胞,胞质从苯丙酸诺龙经处理的hs - 72细胞没有影响在细胞的可行性。concanamycin经处理的hs - 72细胞通过一个四个步骤过程:超速离心法;柱层析法;色谱法;C18液相色谱法疏水的支持。分数,这是用作部分纯化胞质,给一个特定的乐队的蛋白质分子质量33 kDa的钠十二烷基sulfate-polyacrylamide凝胶电泳。细胞死亡机制被观察检查核形态的变化,增加比例的DNA片段,典型的梯子的退化模式染色体DNA,表明细胞凋亡的诱导细胞培养与部分纯化细胞溶质。超表达人类的bcl - 2抑制细胞凋亡,表明细胞溶质concanamycin经处理的hs - 72细胞诱发Bcl-2-inhibiting细胞凋亡的机制。研究结果表明,concanamycin,空泡的型H + atp酶抑制剂,产生胞内B细胞杂种细胞的凋亡诱导因素。

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