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microRNA-141-3p-containing small extracellular vesicles derived from epithelial ovarian cancer cells promote endothelial cell angiogenesis through activating the JAK/STAT3 and NF-κB signaling pathways

Abstract

Paracrine signaling between tumor and surrounding stromal cells is critical for the maintenance of tumor microenvironment during ovarian cancer progression. Small extracellular vesicles (sEVs; exosomes in particular) are nano-sized vesicles secreted actively by many cells including tumor cells and are found to have fundamental roles in intercellular communication through shuttling functional RNAs. Although microRNAs (also called miRNAs or miRs), small non-coding RNAs regulating gene expression, are selectively accumulated in tumor sEVs and can mediate intercellular communication, the exact biological mechanisms underlying the functions of exosomal miRNAs in ovarian tumor angiogenesis remain unclear. In this study, sEVs were isolated from conditioned medium of the human ovarian carcinoma cell line SKOV-3 using ExoQuick Exosome Precipitation Solution, and characterized by scanning electron microscopy, dynamic light scattering, and immunoblotting. To elucidate the possible paracrine effects on ovarian tumor cell-derived sEVs (TD-sEVs), we investigated the angiogenesis-related signaling events triggered by TD-sEVs in endothelial cells. Due to the possible role in ovarian tumor pathogenesis, we focused on miR-141-3p which was detected to be enriched in TD-sEVs compared with their corresponding donor cells. We identified that sEV transfer of miR-141-3p considerably reduced the expression levels of cytokine-inducible suppressors of cytokine signaling (SOCS)-5 leading to up-regulated JAK-STAT3 pathway in endothelial cells. We also observed that sEV-shuttled miR-141-3p may up-regulate the expression of VEGFR-2 in endothelial cells which leads to promoting endothelial cell migration and angiogenesis. The putative role of miR-141-3p shuttled by TD-sEVs in regulating VEGFR-2 expression was demonstrated by the ability of anti-miR-141-3p to rescue the promoting effects of TD-sEVs on the expression of VEGFR-2 in endothelial cells. Our results also revealed that TD-sEVs trigger the intracellular reactive oxygen species (ROS)-dependent activation of NF-κB signaling in endothelial cells. Taken together, our findings propose a novel model in which sEV transfer of epithelial ovarian cancer-secreted miR-141-3p plays as a significant mediator of intercellular communication, promoting endothelial cell angiogenesis.

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Abbreviations

VEGF:

Vascular endothelial growth factor

VEGFR-2:

Vascular endothelial growth factor receptor-2

JAK/STAT:

The Janus kinase/signal transducer and activator of transcription

miRNAs, miRs:

MicroRNAs

sEVs:

Small extracellular vesicles

TD-sEVs:

Ovarian tumor cell-derived small extracellular vesicles

NT-sEVs:

Non-tumorigenic ovarian epithelial cell-derived small extracellular vesicles

NF-κB:

Nuclear factor kappa-light-chain-enhancer of activated B cells

HUVECs:

Human umbilical vein endothelial cells

DLS:

Dynamic light scattering

U6snRNA:

U6 small nuclear RNA

RNU44:

U44 Smal nucleolar RNA

DCFDA:

2′, 7′-dichlorofluorescein diacetate

SEM:

Scanning electron microscopy

ROS:

Reactive oxygen species

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Acknowledgements

We would like to thank Dr. Majid Mossahebi-Mohammadi, Dr. Katayoon Pakravan, and Ms. Fahimeh-Sadet Norouzi for their technical assistance and advice. This work was supported financially by Tarbiat Modares University and Iran National Science Foundation (INSF) (Grant No. 96008647).

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Correspondence to Sadegh Babashah.

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Masoumi-Dehghi, S., Babashah, S. & Sadeghizadeh, M. microRNA-141-3p-containing small extracellular vesicles derived from epithelial ovarian cancer cells promote endothelial cell angiogenesis through activating the JAK/STAT3 and NF-κB signaling pathways. J. Cell Commun. Signal. (2020). https://doi.org/10.1007/s12079-020-00548-5

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Keywords

  • Tumor angiogenesis
  • Small extracellular vesicles
  • Ovarian cancer-secreted miR-141-3p
  • JAK-STAT3 pathway