A proteomic analysis of Bcl-2 regulation of cell cycle arrest: insight into the mechanisms
A proteomic analysis of Bcl-2 regulation of cell cycle arrest: insight into the mechanisms
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DOI：
10.1631/jzus.b2000802
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发表时间：
2021-10-01
影响因子：
5.1
通讯作者：
Pei, Xiuying
中科院分区：
文献类型：
--
作者：
Du, Xing;Xiao, Jingjing;Pei, Xiuying
B细胞淋巴瘤2（Bcl-2）是一种重要的抗凋亡基因，在维持癌细胞生存与死亡的动态平衡中起着双重作用。在我们以前的研究中，Bcl-2被证明通过调节线粒体代谢途径产生较低水平的三磷酸腺苷（ATP）和活性氧（ROS）来延迟G 0/G1到S期的进入。然而，Bcl-2调节细胞周期的详细分子机制或途径仍然未知。在这里，我们比较了Bcl-2过表达与空载体对照在NIH 3 T3细胞系同步血清饥饿的影响，并使用蛋白质组学分析评估的影响。通过检测Bcl-2和p27的表达来检测Bcl-2对细胞周期的调控作用。随后的Bcl-2过表达细胞的蛋白质组学分析的结果鉴定了169个上调和120个下调的蛋白质，变化为1.5倍。根据基因本体论（GO）和京都基因和基因组百科全书（KEGG）富集分析的数据，这些差异表达的蛋白质在许多主要涉及核糖体和氧化磷酸化的信号通路中富集。这些结果表明，Bcl-2可能在翻译水平上影响呼吸链或核糖体中的蛋白质或酶，从而调节细胞周期。此外，参与氧化磷酸化的差异表达蛋白被确定为解释Bcl-2对我们先前研究中研究的线粒体途径介导的细胞周期的大部分影响。这些结果可为进一步深入研究Bcl-2对细胞周期的调控提供帮助。蛋白质组学分析的结果确定了Bcl-2依赖性延迟细胞周期进程的机制。总之，本研究的结果为确定设计和开发精确靶向癌症药物的关键蛋白或途径提供了新的机制基础。
B cell lymphoma 2 (Bcl-2) is an important antiapoptotic gene that plays a dual role in the maintenance of the dynamic balance between the survival and death of cancer cells. In our previous study, Bcl-2 was shown to delay the G0/G1 to S phase entry by regulating the mitochondrial metabolic pathways to produce lower levels of adenosine triphosphate (ATP) and reactive oxygen species (ROS). However, the detailed molecular mechanisms or pathways by which Bcl-2 regulates the cell cycle remain unknown. Here, we compared the effects of Bcl-2 overexpression with an empty vector control in the NIH3T3 cell line synchronized by serum starvation, and evaluated the effects using proteomic analysis. The effect of Bcl-2 on cell cycle regulation was detected by monitoring Bcl-2 and p27 expression. The result of subsequent proteomic analysis of Bcl-2 overexpressing cells identified 169 upregulated and 120 downregulated proteins with a 1.5-fold change. These differentially expressed proteins were enriched in a number of signaling pathways predominantly involving the ribosome and oxidative phosphorylation, according to the data of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. These results indicated that Bcl-2 potentially acts at the translation level to influence proteins or enzymes of the respiratory chain or in the ribosome, and thereby regulates the cell cycle. Additionally, differentially expressed proteins involved in oxidative phosphorylation were determined to account for most of the effects of Bcl-2 on the cell cycle mediated by the mitochondrial pathway investigated in our previous study. These results can provide assistance for additional in-depth studies on the regulation of the cell cycle by Bcl-2. The results of the proteomic analysis determined the mechanism of Bcl-2-dependent delay of the cell cycle progression. In summary, the results of this study provide a novel mechanistic basis for identifying the key proteins or pathways for designing and developing precisely targeted cancer drugs.