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Archives of Pharmacal Research

, Volume 42, Issue 8, pp 684–694 | Cite as

Realgar transforming solution-induced differentiation of NB4 cell by the degradation of PML/RARα partially through the ubiquitin–proteasome pathway

  • Yang Hai
  • Xin Wang
  • Peng Song
  • Jian-yin Li
  • Long-he Zhao
  • Fei Xie
  • Xiang-min Tan
  • Qin-jian Xie
  • Lan Yu
  • Yang Li
  • Zheng-rong Wu
  • Hong-yu LiEmail author
Research Article
  • 124 Downloads

Abstract

PML/retinoic acid receptor alpha (RARα), as a hallmark of acute promyeloid leukemia (APL), is directly related to the outcome of clinical APL remedy. It is reported that arsenicals can effectively degrade PML/RARα, such as arsenic trioxide and realgar. However, the high toxicity or insolubility have hampered their clinical applications. Realgar transforming solution (RTS) was produced from realgar by bioleaching process in our lab. Previous studies demonstrated that RTS had a significant anti-cancer ability on chronic myeloid leukemia through oncoprotein degradation. The capacity of RTS on treating APL is what is focused on in this study. The results showed that RTS had a noticeable sensitivity in NB4 cell, and RTS remarkably down-regulated PML/RARα expression and induced cell differentiation. Further, RTS could accumulate PML/RARα into the nuclear bodies and then execute degradation, which could be reversed by proteasome inhibitor MG132. The results also exhibited that the reduction of RTS-induced PML/RARα expression accompanied by the elevation of ubiquitin and SUMO-1 protein expression. Finally, PML and SUMO-1 had been demonstrated to be co-localized after RTS treatment by immunofluorescence co-localization assay and immunoprecipitation assay. In conclusion, these results suggested that RTS-induced cell differentiation may attribute to the PML/RARα degradation partially through the ubiquitin–proteasome pathway.

Keywords

Realgar Arsenic PML/RARα SUMO Ubiquitin–proteasome pathway 

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant Numbers 81403145, 81560715, 51501080, 81803779), the Sub-Project of National Science and Technology Major Projects for “Major New Drugs Innovation and Development” (Grant Number 2015ZX09501-004-003-008), The Fundamental Research Funds for the Central Universities of China (lzujbky-2018-136, lzujbky-2018-40, lzujbky-2017-206).

Compliance with ethical standards

Conflict of Interest

No potential conflict of interest was reported by the authors.

Supplementary material

12272_2019_1170_MOESM1_ESM.tiff (1 mb)
Supplementary material 1 (TIFF 1073 kb) Supplemental Fig. A Cytotoxicity of NB4 cell in different arsenic agents’ treatments. B The expression level of PML/RAR⍺ after treating with different arsenic agents. C AV/PI analysis for NB4 cell apoptosis of RTS and ATO incubated in or without MG132. D The expression of CD11b after treating with ATO. E Wright–Giemsa staining after treating with RTS and ATO. The solid red arrows indicated the lobulated nuclei, which represented differentiation cells (× 400)

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Copyright information

© The Pharmaceutical Society of Korea 2019

Authors and Affiliations

  1. 1.School of PharmacyLanzhou UniversityLanzhouChina
  2. 2.Institute of Microbiology, School of Life SciencesLanzhou UniversityLanzhouChina
  3. 3.Gansu Corps Hospital of CAPFLanzhouChina

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