Effect of Beclin-1 gene silencing on autophagy and apoptosis of the prostatic hyperplasia epithelial cells

Authors

  • Rongfu Liu School of Medicine, Tongji University https://orcid.org/0000-0003-1479-1379
  • Song Zhang The First Affiliated Hospital of Xiamen University
  • Rui Wan The First Affiliated Hospital of Xiamen University
  • Jiang Deng The First Affiliated Hospital of Xiamen University
  • Wei Fang Tongji University

DOI:

https://doi.org/10.1016/j.clinsp.2022.100076

Keywords:

Prostatic hyperplasia, Beclin-1 gene, Autophagy, Apoptosis

Abstract

Objectives: This study aims to explore the effect of silencing Beclin-1 gene on autophagy and apoptosis of Benign Prostatic Hyperplasia (BPH) (BPH-1) cells under the condition of Androgen Deprivation (AD) and Autophagy Inhibition (AI).

Methods: Control group (BPH-1 group), empty carrier group (sh-RNA-BPH-1 group) and Beclin-1 silenced group (sh-Beclin1-BPH-1 group) were set. The Beclin-1 gene silencing efficiency was detected by RT-PCR and Western blot. Autophagic flux was monitored by GFP-LC3 cleavage assay and cell apoptosis was analyzed by flow cytometry. The protein expression levels of LC3, Caspase-3, PARP-1, Bcl-2, and Bax were detected by Western blot.

Results: The transfection of sh-Beclin-1 obviously down-regulated the expression of Beclin-1 at both mRNA and protein levels. Under the conditions of AD and AI, silencing of Beclin-1 restrained the autophagy of BPH-1 cells, as evidenced by a decreased number of autophagosomes and down-regulation of LC3-II protein (p < 0.001). The results of flow cytometry showed that the apoptotic rate of sh-Beclin-1 group was elevated significantly compared to the other two groups (p < 0.01). Western blot results showed that silencing of Beclin-1 promoted 89 kd fragmentation of PARP-1 (p < 0.001) and Caspase-3 activation (p < 0.01). Moreover, silencing of Beclin-1 resulted in declined Bcl-2 and augmented Bax protein expression in BPH-1 cells (p < 0.01), which ultimately led to a decreased Bcl-2/Bax ratio.

Conclusions: The results indicated that the silencing of Beclin-1 gene hampered autophagy while activating apoptosis in BPH-1 cells. Thus, Beclin-1 may participate in an antagonistic relationship between autophagy and apoptosis in BPH.

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Published

2022-09-08

Issue

Section

Original Articles

How to Cite

Liu, R., Zhang, S., Wan, R., Deng, J., & Fang, W. (2022). Effect of Beclin-1 gene silencing on autophagy and apoptosis of the prostatic hyperplasia epithelial cells. Clinics, 77, 100076. https://doi.org/10.1016/j.clinsp.2022.100076