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STIM1-regulated exosomal EBV-LMP1 empowers endothelial cells with an aggressive phenotype by activating the Akt/ERK pathway in nasopharyngeal carcinoma

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Abstract

Background

Stromal interaction molecule 1 (STIM1)-mediated Ca2+ signaling regulates tumor angiogenesis in nasopharyngeal carcinoma (NPC), an Epstein-Barr virus (EBV)-related human malignancy. However, the mechanism by which STIM1 modulates endothelial functional phenotypes contributing to tumor angiogenesis remains elusive.

Methods

NPC cell-derived exosomes were isolated via differential centrifugation and observed using transmission electron microscopy. Exosome particle sizes were assessed by nanoparticle tracking analysis (NTA). Uptake of exosomes by recipient ECs was detected by fluorescent labeling of the exosomes with PKH26. Tumor angiogenesis-associated profiles were characterized by determining cell proliferation, migration, tubulogenesis and permeability in human umbilical vein endothelial cells (HUVECs). Activation of the Akt/ERK pathway was assessed by detecting the phosphorylation levels using Western blotting. A chick embryo chorioallantoic membrane (CAM) xenograft model was employed to study tumor-associated neovascularization in vivo.

Results

We found that NPC cell-derived exosomes harboring EBV-encoded latent membrane protein 1 (LMP1) promoted proliferation, migration, tubulogenesis and permeability by activating the Akt/ERK pathway in ECs. STIM1 silencing reduced LMP1 enrichment in NPC cell-derived exosomes, thereby reversing its pro-oncogenic effects in an Akt/ERK pathway-dependent manner. Furthermore, STIM1 knockdown in NPC cells blunted tumor-induced vascular network formation and inhibited intra-tumor neovascularization in the chorioallantoic membrane (CAM) xenograft model.

Conclusion

STIM1 regulates tumor angiogenesis by controlling exosomal EBV-LMP1 delivery to ECs in the NPC tumor microenvironment. Blocking exosome-mediated cell-to-cell horizontal transfer of EBV-associated oncogenic signaling molecules may be an effective therapeutic strategy for NPC.

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Data availability

The original data supporting the conclusions of our article will be available from the corresponding authors upon reasonable request.

Abbreviations

STIM1 :

Stromal interaction molecule 1

NPC :

Nasopharyngeal carcinoma

EBV :

Epstein-Barr virus

LMP1 :

Latent membrane protein 1

HNSCC :

Head and neck squamous cell carcinoma

TME :

Tumor microenvironment

ECs :

Endothelial cells

SOCE :

Store-operated Ca2+ entry

CAM :

Chorioallantoic membrane

FBS :

Fetal bovine serum

NTA :

Nanoparticle tracking analysis

HUVECs :

Human umbilical vein endothelial cells

PBS :

Phosphate-buffered saline

CCK-8 :

Cell Counting Kit-8

EdU :

5-Ethynyl-2′-deoxyuridine

FITC :

Fluorescein isothiocyanate

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Acknowledgements

We sincerely thank Prof. Sai-Wah Tsao (Hong Kong University) and Prof. Musheng Zeng (Sun Yat-sen University Cancer Center) for the gift of NPC cell lines.

Funding

This work was supported by the National Natural Science Foundation of China [grant numbers 82002859, 82073004, 81602390]; the Guangxi Natural Science Foundation [grant numbers 2020GXNSFBA297024, 2020GXNSFBA297059]; and Guangxi Medical and Health Key Discipline Construction Project. The sponsors had no role in the study design, data collection/analysis, manuscript writing, or the decision to submit the paper for publication.

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Contributions

J. Wei and J. Zhang conceived and designed the study. Y. Deng, J. Wei, and J. Zhang wrote the manuscript and prepared the figures. Y. Deng, X. Liu, Y. Huang, J. Ye, Y. Luo, Y. Chen, and Q. Li performed the experiments. Y. Deng, Q. He, Y. Lin, R. Liang, Y. Li, J. Wei, and J. Zhang analyzed the results. All the authors reviewed drafts of the paper and approved the submission of the manuscript.

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Correspondence to Jiazhang Wei or Jinyan Zhang.

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This study was approved by the ethics committee of Guangxi Medical University Cancer Hospital (Ref: KY2020014), and informed consent was obtained from all patients. This study complied with the principles of the Declaration of Helsinki. All animal experiments performed in this study were approved by the Animal Research Committee of Guangxi Medical University and carried out according to the regulations of the Animal Research Committee of Guangxi Medical University.

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Deng, Y., Liu, X., Huang, Y. et al. STIM1-regulated exosomal EBV-LMP1 empowers endothelial cells with an aggressive phenotype by activating the Akt/ERK pathway in nasopharyngeal carcinoma. Cell Oncol. 46, 987–1000 (2023). https://doi.org/10.1007/s13402-023-00790-0

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