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FBXO32 targets PHPT1 for ubiquitination to regulate the growth of EGFR mutant lung cancer

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Abstract

Background

Phosphohistidine phosphatase 1 (PHPT1) is an oncogene that has been reported to participate in multiple tumorigenic processes. As yet, however, the role of PHPT1 in lung cancer development remains uncharacterized.

Methods

RNA sequencing assay and 18 pairs of tumor and normal tissues from patients were analyzed to reveal the upregulation of PHPT1 in lung cancer, followed by confirming the biological function in vitro and in vivo. Next, Gene Set Enrichment Analysis, lung cancer samples, apoptosis assay, mass spectrometry experiments and western blotting were used to investigate the molecular mechanism underlying PHPT1 driven progression in epidermal growth factor receptor (EGFR)-mutant lung cancer. Finally, we performed cellular and animal experiments to explore the tumor suppressive function of F-box protein 32 (FBXO32).

Results

We found that PHPT1 is overexpressed in lung cancer patients and correlates with a poor overall survival. In addition, we found that the expression of PHPT1 is elevated in EGFR-mutant lung cancer cells and primary patient samples. Inhibition of PHPT1 expression in EGFR mutant lung cancer cells significantly decreased their proliferation and clonogenicity, and suppressed their in vitro tumor growth. Mechanistic studies revealed that activation of the ERK/MAPK pathway is driven by PHPT1. PHPT1 is required for maintaining drug resistance to erlotinib in EGFR mutant lung cancer cells. We found that FBXO32 acts as an E3 ubiquitin ligase for PHPT1, and that knockdown of FBXO32 leads to PHPT1 accumulation, activation of the ERK/MAPK pathway and promotion of the proliferation, clonogenicity and growth of lung cancer cells.

Conclusions

Our findings indicate that PHPT1 may serve as a biomarker and therapeutic target for acquired erlotinib resistance in lung cancer patients carrying EGFR mutations.

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Availability of data and material

The RNA-sequencing data generated in this research are deposited in Sequence Read Archive (SRA) database. (reviewerlink:https://dataview.ncbi.nlm.nih.gov/object/PRJNA797898?reviewer=sl2j9hkl157fjs3l0avukv9h8q). Survival data supporting this article are from the Kaplan–Meier plotter website and GEO and TCGA datasets, which have been cited.

Code availability

Not applicable.

Abbreviations

PHPT1:

Phosphohistidine phosphatase 1

EGFR:

Epidermal growth factor receptor

FBXO32:

Epidermal growth factor receptor

AAb:

Autoantibody

AKT/mTOR pathway:

Protein kinase B/mammalian target of rapamycin pathway

ERK/MAPK pathway:

Extracellular signal-regulated/mitogen-activated protein kinase pathway

JNK:

C-Jun N-terminal kinase

BMK-1:

Big MAP kinase-1

MAPK:

Mitogen-activated protein kinase

OD value:

Optical density value

BSA:

Bovine serum albumin

TKI:

Tyrosine kinase inhibitor

NSCLC:

Non-small cell lung cancer

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Acknowledgements

We thank the National Natural Science Foundation of China for support.

Funding

This work was supported by the National Natural Science Foundation of China (81972740 to H.Y.Z.) and the Zhuhai Science and Technology Project (20181117E030079 to Y.F.L. and 20171009E030079 to M.X.).

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Author notes

  1. Ning Zhang, Yifeng Liao and Weize Lv contributed equally.

Authors and Affiliations

  1. Internal Medicine Oncology, Minhang Brunch Fudan University Shanghai Cancer Center, Shanghai, 200240, China

    Ning Zhang

  2. Department of Oncology, The Fifth Affiliated Hospital of Sun Yat-Sen University, Zhuhai, 519000, China

    Yifeng Liao, Weize Lv, Yeqing Qiu, Nan Chen, Mei Xiao & Hongyu Zhang

  3. Department of Neurosurgery, Huizhou Third People’s Hospital, Guangzhou Medical University, 516000, Huizhou, China

    Shunda Zhu

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  1. Ning Zhang
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Contributions

N.Z., Y.F.L. and W.Z.L. performed the experiments. Y.Q.Q. and N.C. analyzed and interpreted data. S.D.Z. polished the manuscript. M.X. and H.Y.Z. provided ideas and critical comments. M.X. and H.Y.Z. conceived and designed the study and co-wrote the paper with feedback from all authors. All authors approved the final manuscript.

Corresponding authors

Correspondence to Mei Xiao or Hongyu Zhang.

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The study was performed in accordance with the Declaration of Helsinki. All human specimen and cell studies were reviewed and approved by the Ethics Committee of The Fifth Affiliated Hospital of Sun Yat-sen University, and informed written consent was obtained from all donors.

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This study is compliant with all relevant ethical regulations regarding animal research. Animal experiments were approved by the Animal Ethical and Welfare Research Committee of the Fifth Affiliated Hospital of Sun Yat-sen University and performed in accordance with established ARRIVE guidelines.

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Zhang, N., Liao, Y., Lv, W. et al. FBXO32 targets PHPT1 for ubiquitination to regulate the growth of EGFR mutant lung cancer. Cell Oncol. 45, 293–307 (2022). https://doi.org/10.1007/s13402-022-00669-6

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