MBD2 upregulates MIR-301a-5p to induce kidney cell apoptosis during vancomycin-induced AKI

Juan Wang, Huiling Li, Shuangfa Qiu, Zheng Dong, Xudong Xiang, Dongshan Zhang

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Despite DNA methylation occurred in acute kidney injury (AKI), how it influenced progression of AKI remains unclear. Methyl-CpGbinding domain protein 2 (MBD2), a protein readers of methylation, was used to analyze the impact of DNA methylation on vancomycin (VAN)-induced AKI. Here, in cultured human kidney tubular epithelial cells (HK-2), we show that knockdown of MBD2 by siRNA attenuated VAN-induced apoptosis, caspase activity, and the expression of BAX and cleaved caspase 3. Interestingly, knockdown of MBD2 by siRNA was associated with the suppression of miR-301a-5p. Mechanistic studies confirmed MBD2 binds to these methylated CpG elements of miR-301a-5p promoter, and then activates miR-301a-5p promoter by suppressing methylation. Furthermore, anti-miR-301a-5p significantly blocked VAN-induced apoptosis and caspase activity in HK-2 cells, which was accompanied by downregulation of p53, and upregulation of MITF, HDGF and MDM-4 together. The latter genes were further identified as target genes of miR-301a-5p, and silencing of MDM-4 promoted p53 accumulation. In vivo, mice with MBD2 knockout (MBD2-KO) were counteracted to VAN-induced AKI, indicated by the analysis of renal function, histology, apoptosis and inflammation. MBD2-KO also significantly suppressed the expression of miR-301a-5p, p53, BAX and cleaved caspase 3, and restored the expression of MDM-4, MITF and HDGF. Finally, in vivo inhibition of miR-301a-5p also ameliorated VAN-induced AKI. Together, these results show the novel MBD2/miR-301a-5p/MITF, HDGF and MDM-4/p53 pathway in VAN-induced AKI.

Original languageEnglish (US)
Article numbere3120
JournalCell Death and Disease
Volume8
Issue number10
DOIs
StatePublished - Oct 12 2017

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Vancomycin
Acute Kidney Injury
Up-Regulation
Apoptosis
Kidney
DNA Methylation
Caspases
Caspase 3
Methylation
Small Interfering RNA
Genes
Histology
Down-Regulation
Epithelial Cells
Inflammation

ASJC Scopus subject areas

  • Immunology
  • Cellular and Molecular Neuroscience
  • Cell Biology
  • Cancer Research

Cite this

MBD2 upregulates MIR-301a-5p to induce kidney cell apoptosis during vancomycin-induced AKI. / Wang, Juan; Li, Huiling; Qiu, Shuangfa; Dong, Zheng; Xiang, Xudong; Zhang, Dongshan.

In: Cell Death and Disease, Vol. 8, No. 10, e3120, 12.10.2017.

Research output: Contribution to journalArticle

Wang, Juan ; Li, Huiling ; Qiu, Shuangfa ; Dong, Zheng ; Xiang, Xudong ; Zhang, Dongshan. / MBD2 upregulates MIR-301a-5p to induce kidney cell apoptosis during vancomycin-induced AKI. In: Cell Death and Disease. 2017 ; Vol. 8, No. 10.
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abstract = "Despite DNA methylation occurred in acute kidney injury (AKI), how it influenced progression of AKI remains unclear. Methyl-CpGbinding domain protein 2 (MBD2), a protein readers of methylation, was used to analyze the impact of DNA methylation on vancomycin (VAN)-induced AKI. Here, in cultured human kidney tubular epithelial cells (HK-2), we show that knockdown of MBD2 by siRNA attenuated VAN-induced apoptosis, caspase activity, and the expression of BAX and cleaved caspase 3. Interestingly, knockdown of MBD2 by siRNA was associated with the suppression of miR-301a-5p. Mechanistic studies confirmed MBD2 binds to these methylated CpG elements of miR-301a-5p promoter, and then activates miR-301a-5p promoter by suppressing methylation. Furthermore, anti-miR-301a-5p significantly blocked VAN-induced apoptosis and caspase activity in HK-2 cells, which was accompanied by downregulation of p53, and upregulation of MITF, HDGF and MDM-4 together. The latter genes were further identified as target genes of miR-301a-5p, and silencing of MDM-4 promoted p53 accumulation. In vivo, mice with MBD2 knockout (MBD2-KO) were counteracted to VAN-induced AKI, indicated by the analysis of renal function, histology, apoptosis and inflammation. MBD2-KO also significantly suppressed the expression of miR-301a-5p, p53, BAX and cleaved caspase 3, and restored the expression of MDM-4, MITF and HDGF. Finally, in vivo inhibition of miR-301a-5p also ameliorated VAN-induced AKI. Together, these results show the novel MBD2/miR-301a-5p/MITF, HDGF and MDM-4/p53 pathway in VAN-induced AKI.",
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