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Gastric Cancer
Published in: Gastric Cancer 5/2020

01-09-2020 | Epstein-Barr Virus | Original Article

Epstein–Barr virus-encoded miR-BART5-5p upregulates PD-L1 through PIAS3/pSTAT3 modulation, worsening clinical outcomes of PD-L1-positive gastric carcinomas

Authors: Chan Jin Yoon, Mee Soo Chang, Dong Ha Kim, Won Kim, Bo Kyung Koo, Sung-Cheol Yun, Sung Han Kim, Yang Soo Kim, Jun Hee Woo

Published in: Gastric Cancer | Issue 5/2020

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Abstract

Background

Epstein–Barr virus (EBV) is etiologically associated with ~ 10% of all gastric carcinomas. However, the molecular mechanisms and roles of EBV miRNAs in gastric carcinoma oncogenesis are yet to be elucidated.

Methods

MicroRNA microarray and TaqMan quantitative real-time RT-PCR were conducted. RT-PCR and luciferase reporter assay for PIAS3, western blotting for 20 proteins, immunofluorescence for STAT3, transfection with miRBART5-5p-plasmid, STAT3-plasmid, miRBART5-5p mimic, or PIAS3-siRNA, and in vitro assays for biological effects of PD-L1 were implemented. In situ hybridization for EBV-encoded small RNAs and immunohistochemistry were performed on gastric carcinoma tissues.

Results

Transfecting miR-BART5-5p into EBV(−) gastric carcinoma cell lines caused a decrease in PIAS3 3′-UTR reporter activity, PIAS3 downregulation, and subsequent STAT3 activation followed by PIAS3/pSTAT3-dependent PD-L1 upregulation. Interestingly, due to PD-L1 knockdown, apoptosis was increased, while the rate of cell proliferation, invasion capacity, and migration were decreased in miR-BART5-5p-transfected cells. In EBV(+) gastric carcinoma cells, anti-miR-BART5-5p reduced PD-L1 levels through PIAS3/pSTAT3 control. Among 103 patients with EBV-associated gastric carcinomas, overall survival was significantly shortened for those with PD-L1(+) tumors compared to those with PD-L1(−) tumors (P = 0.049).

Conclusions

Our findings imply that miR-BART5-5p directly targets PIAS3 and augments PD-L1 through miR-BART5/PIAS3/pSTAT3/PD-L1 axis control. This contributes to antiapoptosis, tumor cell proliferation, invasion and migration, as well as immune escape, furthering gastric carcinoma progression and worsening the clinical outcome, especially in the PD-L1(+) group of patients with EBV-associated gastric carcinomas. miR-BART5-5p may, therefore, be amenable to PD-1/PD-L1 immune checkpoint inhibitor therapy.
Appendix
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Metadata
Title
Epstein–Barr virus-encoded miR-BART5-5p upregulates PD-L1 through PIAS3/pSTAT3 modulation, worsening clinical outcomes of PD-L1-positive gastric carcinomas
Authors
Chan Jin Yoon
Mee Soo Chang
Dong Ha Kim
Won Kim
Bo Kyung Koo
Sung-Cheol Yun
Sung Han Kim
Yang Soo Kim
Jun Hee Woo
Publication date
01-09-2020
Publisher
Springer Singapore
Published in
Gastric Cancer / Issue 5/2020
Print ISSN: 1436-3291
Electronic ISSN: 1436-3305
DOI
https://doi.org/10.1007/s10120-020-01059-3

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