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07 June 2020 : Laboratory Research  

Autophagy Inhibition Sensitizes Renal Tubular Epithelial Cell to G1 Arrest Induced by Transforming Growth Factor beta (TGF-β)

Chen Yang1ACEG, Hong-luan Wu1BCDE, Zhi-hang Li1CDEF, Xiao-cui Chen1BCD, Hong-yong Su1BF, Xiao-yan Guo1CD, Ning An1EFG, Kai-peng Jing1EF, Qing-jun Pan1EF, Hua-feng Liu1ADEFG*

DOI: 10.12659/MSM.922673

Med Sci Monit 2020; 26:e922673


BACKGROUND: Cell cycle arrest and autophagy have been demonstrated to be involved in various transforming growth factor (TGF)-β-mediated phenotype alterations of tubular epithelial cells (TECs) and tubulointerstitial fibrosis. But the relationship between cell cycle arrest and the autophagy induced by TGF-β has not been explored well.

MATERIAL AND METHODS: The effects of autophagy inhibition on TGF-β-induced cell cycle arrest in TECs were explored in vitro. Human kidney-2 (HK-2) cells were stimulated by TGF-β with or without a combined treatment of autophagy inhibitor chloroquine (CQ) or bafilomycin A1 (Baf).

RESULTS: Autophagy inhibition by CQ or Baf promotes the suppression of growth in TGF-β-treated HK-2 cells, as detected by the Cell Counting Kit-8 (CCK-8) method. In addition, CQ or Baf stimulation enhances G1 arrest in TGF-β treated HK-2 cells, as investigated using propidium iodide (PI) staining and flow cytometry, which was further confirmed by a decrease in the expression of phosphorylated retinoblastoma protein (p-RB) and cyclin-dependent kinase 4 (CDK4). The upregulation of p21 induced by CQ or Baf may mediate an enhanced G1 arrest in TGF-β treated HK-2 cells. Western blot analysis showed that TGF-β-induced expression of extracellular matrix fibronectin was notably upregulated in the presence of autophagy inhibitors.

CONCLUSIONS: Inhibition of autophagy sensitizes the TECs to G1 arrest and proliferation suppression induced by TGF-β that contributes to the induction of tubulointerstitial fibrosis.

Keywords: Fibrosis, G1 Phase Cell Cycle Checkpoints, Transforming Growth Factor beta, Cell Line, Chloroquine, Cyclin-Dependent Kinase 4, Cyclin-Dependent Kinase Inhibitor p21, Fibronectins, In Vitro Techniques, Kidney Tubules, Macrolides, Renal Insufficiency, Chronic, Retinoblastoma Protein



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Editorial: Global Surveillance of Highly Pathogenic Avian Influenza Viruses in Poultry, Wild Birds, and Mammals to Prevent a Human Influenza Pandemic

Dinah V. Parums
Science Editor, Medical Science Monitor, International Scientific Information, Inc., Melville, NY, USA

DOI: 10.12659/MSM.939968

Med Sci Monit 2023; 29:e939968


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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750