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首页> 外文学位 >Role of Bcl-2 proteins in stress-induced apoptosis in a non-transformed mammary epithelial cell line.
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Role of Bcl-2 proteins in stress-induced apoptosis in a non-transformed mammary epithelial cell line.

机译：Bcl-2蛋白在非转化乳腺上皮细胞系中在应激诱导的细胞凋亡中的作用。

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摘要

Apoptosis is the process of programmed cell death characterized by morphological and physiological changes executed by cells in response to a stimulus. The apoptotic process contributes to the loss of milk secreting cells (i.e. mammary epithelial cells) that occurs in the bovine mammary gland once peak lactation has occurred. However, the specific mechanisms by which this occurs are unknown. The goal of this work was to examine the role of the Bcl-2 family of proteins in regulating apoptosis in the bovine mammary epithelial cell line, MAC-T.;The Bcl-2 family is divided into three categories based on protein function and combinations of four Bcl-2 homology (BH) domains. Anti-apoptotic multidomain proteins, such as Bcl-2 and Mcl-1, prevent apoptosis by binding to pro-apoptotic members of the family. Pro-apoptotic multidomain proteins, such as Bax, help to regulate apoptosis at the mitochondrial outer membrane. Pro-apoptotic BH3-only proteins, such as Bad and Bim, act as messengers between the anti-apoptotic and pro-apoptotic proteins.;To investigate the roles of Bcl-2 proteins in apoptosis, MAC-T cells were treated with anisomycin (ANS), an activator of the intrinsic apoptotic pathway. ANS had little effect on Bcl-2 or Bax mRNA levels while it induced a 40 to 60% decrease in levels of Bad mRNA. Protein expression of Bax did not change during apoptosis, while Bcl-2, Mcl-1, and Bad expression decreased to varying degrees. The greatest change in protein levels was observed for Mcl-1, whose expression was nearly non-detectable after 4 h of treatment with ANS. Bim was phosphorylated in response to ANS but this was not caused by JNK or p38 signaling. Knock-down of Bim using siRNA decreased the ability of ANS to induce apoptosis. Mcl-1 and Bim protein expression changed in a similar time frame, therefore, interactions between the two proteins were evaluated using co-immunoprecipitation experiments. Mcl-1 and Bim interacted both basally and after treatment with ANS. The significance of Bim phosphorylation, the kinase responsible for its phosphorylation, and the functional significance of the interaction between Bim and Mcl-1 in stress-induced apoptosis remain to be determined.
机译：凋亡是程序性细胞死亡的过程，其特征在于细胞响应刺激而执行的形态和生理变化。一旦发生高峰泌乳，凋亡过程会导致牛乳腺中乳汁分泌细胞（即乳腺上皮细胞）的损失。但是，发生这种情况的具体机制尚不清楚。这项工作的目的是检查蛋白Bcl-2家族在调节牛乳腺上皮细胞系MAC-T凋亡中的作用;根据蛋白功能和组合将Bcl-2家族分为三类的四个Bcl-2同源性（BH）域中。抗凋亡的多域蛋白，例如Bcl-2和Mcl-1，通过与该家族的促凋亡成员结合来防止凋亡。促凋亡的多域蛋白（例如Bax）有助于调节线粒体外膜的凋亡。 Bad和Bim等促凋亡的仅BH3蛋白充当抗凋亡蛋白和促凋亡蛋白之间的信使。;为研究Bcl-2蛋白在凋亡中的作用，用茴香霉素处理了MAC-T细胞（ ANS），内在凋亡途径的激活剂。 ANS对Bcl-2或Bax mRNA水平几乎没有影响，而引起Bad mRNA水平降低40％至60％。在凋亡过程中，Bax的蛋白表达没有变化，而Bcl-2，Mcl-1和Bad的表达则有不同程度的下降。对于Mcl-1，观察到蛋白质水平的最大变化，在用ANS处理4小时后其表达几乎不可检测。 Bim响应ANS而被磷酸化，但这不是由JNK或p38信号传导引起的。使用siRNA抑制Bim降低了ANS诱导凋亡的能力。 Mcl-1和Bim蛋白表达在相似的时间范围内发生变化，因此，使用共免疫沉淀实验评估了这两种蛋白之间的相互作用。 Mcl-1和Bim基本相互作用，并在ANS治疗后相互作用。 Bim磷酸化的重要性，负责其磷酸化的激酶，以及Bim和Mcl-1之间的相互作用在应激诱导的细胞凋亡中的功能意义尚待确定。

著录项

  • 作者

    Butler, Kristie Lynn.;

  • 作者单位

    Rutgers The State University of New Jersey - New Brunswick and University of Medicine and Dentistry of New Jersey.;

  • 授予单位 Rutgers The State University of New Jersey - New Brunswick and University of Medicine and Dentistry of New Jersey.;
  • 学科 Cellular biology.;Physiology.;Molecular biology.
  • 学位 M.S.
  • 年度 2014
  • 页码 67 p.
  • 总页数 67
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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