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PHLDA1 is a P53 target gene involved in P53-mediated cell apoptosis

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Abstract

Pleckstrin homeolike domain, family A, member 1 (PHLDA1) is a multifunctional protein that plays diverse roles in A variety of biological processes, including cell death, and hence its altered expression has been found in different types of cancer. Although studies have shown a regulatory relationship between p53 and PHLDA1, the molecular mechanism is still unclear. Especially, the role of PHLDA1 in the process of apoptosis is still controversial. In this study, we found that the expression of PHLDA1 in human cervical cancer cell lines was correlated with the up-expression of p53 after treatment with apoptosis-inducing factors. Subsequently, the binding site and the binding effect of p53 on the promoter region of PHLDA1 were verified by our bioinformatics data analysis and luciferase reporter assay. Indeed, we used CRISPR-Cas9 to knockout the p53 gene in HeLa cells and further confirmed that p53 can bind to the promoter region of PHLDA1 gene, and then directly regulate the expression of PHLDA1 by recruiting P300 and CBP to change the acetylation and methylation levels in the promoter region. Finally, a series of gain–of–function experiments further confirmed that p53 re-expression in HeLap53-/- cell can up-regulate the reduction of PHLDA1 caused by p53 knockout, and affect cell apoptosis and proliferation. Our study is the first to explore the regulatory mechanism of p53 on PHLDA1 by using the p53 gene knockout cell model, which further proves that PHLDA1 is a target-gene in p53-mediated apoptosis, and reveals the important role of PHLDA1 in cell fate determination.

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Data availability

The original contributions presented in the study are included in the article/Supplementary Material. Further inquiries can be directed to the corresponding authors.

Abbreviations

PHLDA1:

Pleckstrin homology-like domain, family A, member 1

DMSO:

Dimethyl sulfoxide

FBS:

Fetal bovine serum

UV:

Ultra violet irradiation

5-FU:

5-Fluorouracil

ChIP:

Chromatin immunoprecipitation assay

OE:

Over expression

SEM:

Standard error of the mean

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Funding

This work was supported by the Guangdong Natural Science Foundation (2020A1515010174), the Distinctive Innovation Grant of Guangdong Department of Education (2020KTSCX035), and Provincial Natural Science Foundation cultivation special project (210719166884337, 210719166884340).

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Authors and Affiliations

  1. Center for Cancer Research, Shantou University Medical College, Shantou, Guangdong, China

    Xuhong Song

  2. Section of Cell Biology and Genetics, Shantou University Medical College, Shantou, Guangdong, China

    Xuhong Song, Lulu Zhou, Wenrui Yang, **nyan Li, Jiazi Ma, Kun Qi, Rui Liang, Mei**g Li, Lingzhu **e, Tin Su, Dongyang Huang & Bin Liang

  3. Biomedical Research Center, Shantou University Medical College, Shantou, Guangdong, China

    Bin Liang

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  1. Xuhong Song
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  2. Lulu Zhou
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Contributions

XS, DH and BL conceived the research and designed the experiments. LZ, WY, XL, JM, KQ, RL, ML, LX and TS performed the experiments. BL wrote the manuscript. BL, XS and DH analyzed the data. All authors revised the manuscript and approved its final version.

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Correspondence to Dongyang Huang or Bin Liang.

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Song, X., Zhou, L., Yang, W. et al. PHLDA1 is a P53 target gene involved in P53-mediated cell apoptosis. Mol Cell Biochem 479, 653–664 (2024). https://doi.org/10.1007/s11010-023-04752-w

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  • DOI: https://doi.org/10.1007/s11010-023-04752-w

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