Effects of bufalin on up-regulating methylation of Wilm’s tumor 1 gene in human erythroid leukemic cells

Li-pei Wang; Yan-na Zhao; Xin Sun; Rui-lan Gao

doi:10.1007/s11655-017-2404-1

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Effects of bufalin on up-regulating methylation of Wilm’s tumor 1 gene in human erythroid leukemic cells

Updated：2021-08-27

- Effects of bufalin on up-regulating methylation of Wilm’s tumor 1 gene in human erythroid leukemic cells
- Chinese Journal of Integrative Medicine Vol. 23, Issue 4, Pages: 288-294(2017)
- Affiliations：
  
  1. College of Basic Medical Sciences, Zhejiang Chinese Medical University,Hangzhou,China
  2. Institution of Oncology, Zhejiang Provincial People’s Hospital,Hangzhou,China
- Author bio：
- Funds：
  
  Supported by National Natural Science Foundation of China (No. 81403223), Zhejiang Provincial Natural Science Foundation of China (No. LQ14H290003) and Science and Technology Foundation of Zhejiang Province (No. 2015C33173)
- DOI：10.1007/s11655-017-2404-1
  CLC：
- Published：2017，
  
  Published Online：1 April 2017，
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Wang, Lp., Zhao, Yn., Sun, X. et al. Effects of bufalin on up-regulating methylation of Wilm’s tumor 1 gene in human erythroid leukemic cells., Chin. J. Integr. Med. 23, 288–294 (2017). https://doi.org/10.1007/s11655-017-2404-1

Li-pei Wang, Yan-na Zhao, Xin Sun, et al. Effects of bufalin on up-regulating methylation of Wilm’s tumor 1 gene in human erythroid leukemic cells. [J]. Chinese Journal of Integrative Medicine 23(4):288-294(2017)
Wang, Lp., Zhao, Yn., Sun, X. et al. Effects of bufalin on up-regulating methylation of Wilm’s tumor 1 gene in human erythroid leukemic cells., Chin. J. Integr. Med. 23, 288–294 (2017). https://doi.org/10.1007/s11655-017-2404-1 DOI：

Li-pei Wang, Yan-na Zhao, Xin Sun, et al. Effects of bufalin on up-regulating methylation of Wilm’s tumor 1 gene in human erythroid leukemic cells. [J]. Chinese Journal of Integrative Medicine 23(4):288-294(2017) DOI： 10.1007/s11655-017-2404-1.

摘要

To explore the effects of bufalin on inhibiting proliferation

up-regulating methylation of Wilm’ tumor 1 gene (WT1) as well as its possible mechanisms in human erythroid leukemic (HEL) cells. The HEL cells were treated with bufalin at various concentrations to observe cellular morphology

proliferation assay and cell cycle. The mRNA and protein expression levels of WT1 were detected by reverse transcription polymerase chain reaction (RT-PCR)

Western blot and immunocytochemistry

DNA methylation of WT1 and protein expression levels of DNA methyltransferase 3a (DNMT3a) and DNMT3b were analyzed by methylation-specific PCR

and Western blot respectively. The bufalin was effective to inhibit proliferation of HEL cells in a dose-dependent manner

their suppression rates were from 23.4%±2.1% to 87.2%±5.4% with an half maximal inhibit concentration (IC50) of 0.046 μmol/L. Typical apoptosis morphology was observed in bufalin-treated HEL cells. The proliferation index of cell cycle decreased from 76.4%±1.9% to 49.7%±1.3%. The expression levels of WT1 mRNA and its protein reduced gradually with increasing doses of bufalin

meanwhile

the methylation status of WT1 gene changed from unmethylated into partially or totally methylated. While

the expression levels of DNMT3a and DNMT3b protein gradually increased by bufalin treatment in a dose-dependent manner. Bufalin can not only significantly inhibit the proliferation of HEL cells and arrest cell cycle at G0/G1 phase

but also induce cellular apoptosis and down-regulate the expression level of WT1. Our results provide the evidence of bufalin for anti-leukemia

its mechanism may involve in increasing WT1 methylation status which is related to the up-regulation of DNMT3a and DNMT3b proteins in erythroid leukemic HEL cells.

Abstract

To explore the effects of bufalin on inhibiting proliferation

proliferation assay and cell cycle. The mRNA and protein expression levels of WT1 were detected by reverse transcription polymerase chain reaction (RT-PCR)

Western blot and immunocytochemistry

DNA methylation of WT1 and protein expression levels of DNA methyltransferase 3a (DNMT3a) and DNMT3b were analyzed by methylation-specific PCR

and Western blot respectively. The bufalin was effective to inhibit proliferation of HEL cells in a dose-dependent manner

meanwhile

the methylation status of WT1 gene changed from unmethylated into partially or totally methylated. While

but also induce cellular apoptosis and down-regulate the expression level of WT1. Our results provide the evidence of bufalin for anti-leukemia

its mechanism may involve in increasing WT1 methylation status which is related to the up-regulation of DNMT3a and DNMT3b proteins in erythroid leukemic HEL cells.

关键词

bufalinhuman erythroid leukemic cellsWilm’s tumor 1 genemethylationDNA methyltransferase 3aDNA methyltransferase 3b

Keywords

bufalinhuman erythroid leukemic cellsWilm’s tumor 1 genemethylationDNA methyltransferase 3aDNA methyltransferase 3b

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Research Center for Traditional Chinese Medicine Complexity System, Shanghai University of Traditional Chinese Medicine

Department of Hematology, Affiliated Hospital of Shandong University of Traditional Chinese Medicine

Postdoctoral Research Station, Shandong University

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Institute of Geriatric Medicine, Xiyuan Hospital, China Academy of Chinese Medical Sciences

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