Scholarlink

Abstract <p><b>Objective:</b> The aim of the work was to find out whether autophagy is activated during the process of replicative aging of cells, and if the activation is detected, study its molecular mechanisms. <b>Methods:</b> The object was the cell line of human eyelid skin dermal fibroblasts DF-2 of mesenchymal origin. The distribution of cells by cell cycle phases, as well as cell size, was determined by flow cytometry. The amount of Atg5, p62, pAMPK, p-4E-BP, LC3-II proteins synthesized by cells was assessed by immunoblotting. The number of autophagic vacuoles was assessed using the CYTO-ID kit and electron microscopy. The development of the senescent phenotype of cells was determined by analyzing the activity of β-galactosidase (SA-β-gal). <b>Results:</b> It was found that aging of human DF-2 fibroblasts was accompanied by activation of autophagy. This type of autophagy is neither cytotoxic nor cytoprotective. Old DF-2 cells were found to increase in size and slow down the cell cycle. In such cells, the Atg5 protein accumulated, the content of p62 protein decreased, the accumulation of the phosphorylated form of AMPK increased, and the synthesis of p-4E-BP1 protein decreased, which indicates low activity of the mTORC1 kinase complex. In addition, the membrane form of the LC3-II protein and autophagic vacuoles accumulate in senescent cells. <b>Conclusion:</b> The aging process of DF-2 cells was accompanied by activation of autophagy, which may be necessary for reprogramming normal cells to a senescent phenotype.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Replicative Aging of Human DF-2 Fibroblasts is Accompanied by the Development of Reprogramming Autophagy

  • 期刊论文

  • 发表日期：

    2025年9月30日

查看全文
  • S. G. Zubova,
  • A. N. Kukushkin,
  • M. G. Martynova,
  • O. A. Bystrova,
  • I. V. Guzhova,
  • B. A. Margulis

摘要

Abstract

Objective: The aim of the work was to find out whether autophagy is activated during the process of replicative aging of cells, and if the activation is detected, study its molecular mechanisms. Methods: The object was the cell line of human eyelid skin dermal fibroblasts DF-2 of mesenchymal origin. The distribution of cells by cell cycle phases, as well as cell size, was determined by flow cytometry. The amount of Atg5, p62, pAMPK, p-4E-BP, LC3-II proteins synthesized by cells was assessed by immunoblotting. The number of autophagic vacuoles was assessed using the CYTO-ID kit and electron microscopy. The development of the senescent phenotype of cells was determined by analyzing the activity of β-galactosidase (SA-β-gal). Results: It was found that aging of human DF-2 fibroblasts was accompanied by activation of autophagy. This type of autophagy is neither cytotoxic nor cytoprotective. Old DF-2 cells were found to increase in size and slow down the cell cycle. In such cells, the Atg5 protein accumulated, the content of p62 protein decreased, the accumulation of the phosphorylated form of AMPK increased, and the synthesis of p-4E-BP1 protein decreased, which indicates low activity of the mTORC1 kinase complex. In addition, the membrane form of the LC3-II protein and autophagic vacuoles accumulate in senescent cells. Conclusion: The aging process of DF-2 cells was accompanied by activation of autophagy, which may be necessary for reprogramming normal cells to a senescent phenotype.