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Effect of Phosphodiesterase in Regulating the Activity of Lysosomes in the HeLa Cell Line

  • Hong, Eun-Seon (Graduate School of Semiconductor and Chemical Engineering, Chonbuk National University) ;
  • Kim, Bit-Na (Graduate School of Semiconductor and Chemical Engineering, Chonbuk National University) ;
  • Kim, Yang-Hoon (Department of Microbiology, Chungbuk National University) ;
  • Min, Jiho (Graduate School of Semiconductor and Chemical Engineering, Chonbuk National University)
  • Received : 2016.07.21
  • Accepted : 2016.10.31
  • Published : 2017.02.28

Abstract

The transport of lysosomal enzymes into the lysosomes depends on the phosphorylation of their chains and the binding of the phosphorylated residues to mannose-6-phosphate receptors. The efficiency of separation depends more on the phosphodiesterases (PDEs) than on the activity of the phosphorylation of mannose residues and can be determined in vitro. PDEs play important roles in regulation of the activation of lysosomes. The expression of proteins was confirmed by western blotting. All PDE4 series protein expression was reduced in high concentrations of rolipram. As a result of observing the fluorescence intensity after rolipram treatment, the lysosomal enzyme was activated at low concentrations and suppressed at high concentrations. High concentrations of rolipram recovered the original function. Antimicrobial activity was not shown in either 10 or $100{\mu}M$ concentrations of rolipram in treated HeLa cells in vitro. However, the higher anticancer activity at lower rolipram concentration was shown in lysosomal enzyme treated with $10{\mu}M$ of rolipram. The anticancer activity was confirmed through cathepsin B and D assay. Tranfection allowed examination of the relationship between PDE4 and lysosomal activity in more detail. Protein expression was confirmed to be reduced. Fluorescence intensity showed decreased activity of lysosomes and ROS in cells transfected with the antisense sequences of PDE4 A, B, C, and D. PDE4A showed anticancer activity, whereas lysosome from cells transfected with the antisense sequences of PDE4 B, C, and D had decreased anticancer activity. These results showed the PDE4 A, B, C, and D are conjunctly related with lysosomal activity.

Keywords

References

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