Interference of Hsp27 Results in Apoptosis Induced by Photodamage via Regulation of Subcellular Localization of p21 in Immortalized Human Keratinocytes

Med Sci Monit. 2019 Oct 8:25:7557-7566. doi: 10.12659/MSM.917164.

Abstract

BACKGROUND Owing to the increased incidence of photodermatosis caused by ultraviolet light in recent years, it is necessary to clarify the mechanisms potential photodamage to the skin and reveal possible therapeutic targets. Heat shock protein 27 (Hsp27) is well known for suppressing apoptosis. The aim of present study was to elucidate possible photoprotective mechanism between Hsp27 and p21 on ultraviolet B (UVB)-induced photodamage. MATERIAL AND METHODS The Hsp27 gene was interfered to assess the expression of its downstream effectors, cell apoptosis, and cell proliferation ability. The cell apoptosis was tested using flow cytometry method. The cell proliferation ability was tested using Cell Counting Kit-8 (CCK-8) assay. The expression of protein was tested using western-blotting method. The expression of mRNA was detected using quantitative reverse transcription polymerase chain reaction (qRT-PCR). The subcellular localization was elucidated using immunofluorescence. RESULTS Hsp27 knockdown decreased cell viability and increased the incidence of UVB-induced apoptosis. Compared with control group, activation of phosphorylated-Akt (p-Akt)-dependent pathway resulted in the nuclear accumulation of p21 and suppression of cell proliferation, while promoting apoptosis in Hsp27 knockdown group. In addition, Hsp27 knockdown increased p53 expression and the Bax: Bcl-2 ratio, which further accelerated the apoptotic process. CONCLUSIONS These findings complemented the mechanism of skin photodamage and demonstrated the photoprotective mechanisms of Hsp27 in HaCaT cells, which might implicate a potential therapeutic target of photodamage and photodermatosis.

MeSH terms

  • Apoptosis / radiation effects
  • Apoptosis Regulatory Proteins / metabolism
  • Cell Proliferation / radiation effects
  • Cell Survival / radiation effects
  • Cells, Cultured
  • Cyclin-Dependent Kinase Inhibitor p21 / metabolism*
  • Flow Cytometry
  • Heat-Shock Proteins / metabolism*
  • Humans
  • Keratinocytes / metabolism*
  • Keratinocytes / pathology
  • Keratinocytes / radiation effects*
  • Molecular Chaperones / metabolism*
  • RNA, Small Interfering / genetics
  • Signal Transduction
  • Skin / metabolism
  • Skin / pathology
  • Skin / radiation effects
  • Tumor Suppressor Protein p53 / metabolism
  • Ultraviolet Rays

Substances

  • Apoptosis Regulatory Proteins
  • CDKN1A protein, human
  • Cyclin-Dependent Kinase Inhibitor p21
  • HSPB1 protein, human
  • Heat-Shock Proteins
  • Molecular Chaperones
  • RNA, Small Interfering
  • TP53 protein, human
  • Tumor Suppressor Protein p53