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Regulations of Reversal of Senescence by PKC Isozymes in Response to 12-O-Tetradecanoylphorbol-13-Acetate via Nuclear Translocation of pErk1/2

  • Lee, Yun Yeong (Department of Biochemistry and Molecular Biology, Ajou University School of Medicine) ;
  • Ryu, Min Sook (Department of Biochemistry and Molecular Biology, Ajou University School of Medicine) ;
  • Kim, Hong Seok (Department of Molecular Medicine, Inha University, College of Medicine) ;
  • Suganuma, Masami (Research Institute for Clinical Oncology, Saitama Cancer Center) ;
  • Song, Kye Yong (Department of Pathology, Chung-Ang University College of Medicine) ;
  • Lim, In Kyoung (Department of Biochemistry and Molecular Biology, Ajou University School of Medicine)
  • Received : 2015.12.24
  • Accepted : 2015.12.31
  • Published : 2016.03.31

Abstract

The mechanism by which 12-O-tetradecanoylphorbol-13-acetate (TPA) bypasses cellular senescence was investigated using human diploid fibroblast (HDF) cell replicative senescence as a model. Upon TPA treatment, protein kinase C (PKC) ${\alpha}$ and $PKC{\beta}1$ exerted differential effects on the nuclear translocation of cytoplasmic pErk1/2, a protein which maintains senescence. $PKC{\alpha}$ accompanied pErk1/2 to the nucleus after freeing it from $PEA-15pS^{104}$ via $PKC{\beta}1$ and then was rapidly ubiquitinated and degraded within the nucleus. Mitogen-activated protein kinase docking motif and kinase activity of $PKC{\alpha}$ were both required for pErk1/2 transport to the nucleus. Repetitive exposure of mouse skin to TPA downregulated $PKC{\alpha}$ expression and increased epidermal and hair follicle cell proliferation. Thus, $PKC{\alpha}$ downregulation is accompanied by in vivo cell proliferation, as evidenced in 7, 12-dimethylbenz(a)anthracene (DMBA)-TPA-mediated carcinogenesis. The ability of TPA to reverse senescence was further demonstrated in old HDF cells using RNA-sequencing analyses in which TPA-induced nuclear $PKC{\alpha}$ degradation freed nuclear pErk1/2 to induce cell proliferation and facilitated the recovery of mitochondrial energy metabolism. Our data indicate that TPA-induced senescence reversal and carcinogenesis promotion share the same molecular pathway. Loss of $PKC{\alpha}$ expression following TPA treatment reduces pErk1/2-activated SP1 biding to the $p21^{WAF1}$ gene promoter, thus preventing senescence onset and overcoming G1/S cell cycle arrest in senescent cells.

Acknowledgement

Supported by : National Research Foundation (NRF)

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