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NRP1 contributes to stemness and potentiates radioresistance via WTAP-mediated m6A methylation of Bcl-2 mRNA in breast cancer

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Abstract

NRP1 is a transmembrane glycoprotein that is highly expressed in a variety of tumors. There is evidence that NRP1 can enhance the stem cell properties of tumor cells, which are thought to be resistant to radiotherapy. This study aims to elucidate the potential mechanism of NRP1 in radiation resistance. We transfected NRP1 siRNA and plasmid in breast cancer cells to detect the expression of cancer stem cell markers by western blot and qRT-PCR. The effect of NRP1 on radiotherapy resistance was assesses by immunofluorescence and flow cytometry. In vivo, we established xenograft tumor model treating with shRNA-NRP1 to assess radiotherapy sensitivity. We found that NRP1 could enhance the stem cell properties and confer radioresistance of breast cancer cells. Mechanistically, we proved that NRP1 reduced IR-induced apoptosis by downregulation of Bcl-2 via methyltransferase WTAP in m6A-depentent way. It is suggested that these molecules may be the therapeutic targets for improving the efficacy of radiotherapy for breast cancer.

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Date availability

The data and materials in the current study are available from the corresponding author on reasonable request.

Abbreviations

BC:

Breast cancer

NRP1:

Neuropilin-1

CSCs:

Cancer stem cells

BCSCs:

Breast cancer stem cells

WTAP:

Wilms tumoe1-associated protein

IHC:

Immunohistochemistry

FS:

Field size

SSD:

Source-surfaced distance

IP:

Immunoprecipitation

OS:

Overall survival

DFS:

Disease-free survival

dsDNA:

Double-stand DNA

MeRIP:

Methylated RNA immunoprecipitation

Bcl-2:

B-cell lymphoma 2

DEPC:

Diethyl pyrocarbonate

EG00229:

Neuropilin 1 (NRP1) receptor antagonist

ER:

Estrogen receptor

EDTA:

Ethylene diamine tetraacetic acid

H&E:

Hematozlin and eosin stain

IR:

Ionizing radiation

m6A:

N6-methyladenosine

PMSF:

Phenylmethanesulfonyl fluoride

RTK:

Receptor tyrosine kinase

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Acknowledgement

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Funding

This work is supported by Project of Jiangsu Health Commission(M2020072).

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Author notes

  1. Yang Wang and Lin Zhang have contributed equally to this work.

Authors and Affiliations

  1. Department of Pathology, Xuzhou Medical University, Xuzhou, 221004, China

    Yang Wang, Lin Zhang, Xiao-Lin Sun & Dong-Sheng Pei

  2. Department of Oncological Radiotherapy, The Second Affiliated Hospital of Xuzhou Medical University, Xuzhou, China

    Ya-Chun Lu, Si Chen & Lan-Sheng Zhang

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  1. Yang Wang
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  2. Lin Zhang
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Contributions

DSP and LSZ: designed the study. YW, LZ: performed the main experiments, and XLS: participated in some experiments. YW: drafted the manuscript, YCL and SC: revised the manuscript. All authors provided final approval and agree to be accountable for all aspects of the work.

Corresponding author

Correspondence to Lan-Sheng Zhang.

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Conflict of interest

The authors declare that they have no competing of interest.

Ethical approval

The experiments were approved by the ethics committee of the Second Affiliated Hospital of Xuzhou Medical University, and The Institutional Animal Care and Use Committee of Xuzhou Medical University. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Wang, Y., Zhang, L., Sun, XL. et al. NRP1 contributes to stemness and potentiates radioresistance via WTAP-mediated m6A methylation of Bcl-2 mRNA in breast cancer. Apoptosis 28, 233–246 (2023). https://doi.org/10.1007/s10495-022-01784-3

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  • DOI: https://doi.org/10.1007/s10495-022-01784-3

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