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• EBV can upregulate the PI3K/AKT pathway through its encoded EBER1 and LMP2A, thereby promoting the expression of YAP1 in EBVaGC cells. • YAP1 enhances the proliferation, migration, autophagy, S-phase accumulation, and anti-apoptotic ability of EBVaGC cells, which may play carcinogenic role in EBVaGC progression. • YAP1 also enhances EBV replication, and TPA and NaB induced EBV lytic reactivation, forming a bidirectional regulatory mechanism with EBV, further promoting the development of EBVaGC. This study aims to explore the role and regulatory mechanism of Yes-associated protein 1 (YAP1) in the development of Epstein-Barr virus-associated gastric cancer (EBVaGC). Here we showed that EBV can upregulate the expression and activity of YAP1 protein through its encoded latent products EBV-encoded small RNA 1 (EBER1) and latent membrane protein 2A (LMP2A), enhancing the malignant characteristics of EBVaGC cells. In addition, we also showed that overexpression of YAP1 induced the expression of EBV encoding latent and lytic phase genes and proteins in the epithelial cell line AGS-EBV infected with EBV, and increased the copy number of the EBV genome, while loss of YAP1 expression reduced the aforementioned indicators. Moreover, we found that YAP1 enhanced EBV lytic reactivation induced by two known activators, 12-O-tetradecanoylhorbol-13-acetate (TPA) and sodium butyrate (NaB). These results indicated a bidirectional regulatory mechanism between EBV and YAP1 proteins, providing new experimental evidence for further understanding the regulation of EBV infection patterns and carcinogenic mechanisms in gastric cancer. [ABSTRACT FROM AUTHOR]

Highlights • The relative expression of miR-BART16 is higher in EBVaGC tissues than cell lines. • Overexpression of miR-BART16 could reduce the LMP1 mRNA and protein levels. • Inhibition the endogenous miR-BART16 increases the LMP1 mRNA and protein level. • BART16 has a vital role on cell proliferation in EBV-associated gastric carcinoma. Abstract Epstein–Barr virus (EBV) is a ubiquitous oncogenic herpesvirus associated with various human tumors. Latent membrane protein (LMP) 1 is considered a key oncoprotein in some EBV-associated tumors, while it is absent in EBV-associated gastric carcinoma (EBVaGC). In the present study, we did not detect the mRNA or the protein expression of LMP1 in 10 EBVaGC tissue samples, but we found that miRNA-BART16 was more abundantly expressed in EBVaGC primary tissues than in cell lines. It has been reported that EBV-encoded miRNA-BART16 can target the 3′-untranslated regions of LMP1 and negatively regulate its expression. When a BART16 inhibitor or mimics were transfected into EBVaGC cell lines, both mRNA and protein levels were changed compared with those in controls. We further investigated the role of miRNA-BART16 in EBVaGC. According to the CCK8 assay, the proliferation of GT38 and GT39 cells increased after treatment with a miRNA-BART16 inhibitor. At the same time, the BART16 inhibitor caused increased accumulation of G2/M phase cells by bromodeoxyuridine and propidium iodide staining analysis. In addition, no obvious difference was observed in apoptosis after treatment with an inhibitor or mimics using annexin V staining. Our findings highlight the viral role of EBV-encoded miRNA-BART16 in regulating oncogenic LMP1 expression and a negative association with the proliferation of EBVaGC. [ABSTRACT FROM AUTHOR]

• Epstein-Barr Virus (EBV) infection may inhibit aryl hydrocarbon receptor (AHR) expression in EBVaGC. • The proliferation and migration effects of AHR pathway was weakened in EBV-infected GC cells. • The expression of AHR in GC cells may be suppressed by the activation of the ERK pathway mediated by EBV-encoded LMP2A. To investigate the function of the aryl hydrocarbon receptor (AHR) pathway in Epstein-Barr Virus (EBV)-associated gastric cancer (EBVaGC) and to explore the relationship between EBV and AHR expression. The expression of AHR in EBVaGC and EBV negative GC (EBVnGC) tissues was detected by immunohistochemistry (IHC). Real-time qPCR (RT-qPCR) and Western blot analysis were used to examine the expression of AHR, cytochrome P450 1A1 (CYP1A1), and cytochrome P450 1B1 (CYP1B1) in gastric cancer cells. The cell proliferation and migration assay were tested by CCK8 and transwell analysis. EBV-encoded latent membrane protein 2A (LMP2A) was over-expressed in SGC7901 cells and silenced in AGS-EBV cells to further identify its role in EBV positive GC cells. It was found that EBV infection inhibited the expression of AHR in gastric cancer tissues and cell lines. We also found that the activation of AHR pathway can promote cell proliferation and migration. However, the function was restricted in EBVaGC cell lines compared with EBVnGC. LMP2A can suppress AHR expression and pathway activation by activating phosphorylation of extracellular signal-regulated kinase (ERK) in EBV positive GC cell lines. EBV-encoded LMP2A regulated the function of the AHR pathway by activating the ERK signal pathway in EBV positive GC cell lines. [ABSTRACT FROM AUTHOR]

• LMP1 and LMP2A inhibit the expression of COX-2 in EBVaGC. • TRAF2 is involved in the inhibitory effect of LMP1/2A on COX-2. • LMP1 also down-regulates COX-2 by reducing phosphorylation of ERK. Epstein-Barr virus-associated gastric cancer (EBVaGC) accounts for nearly 10% of gastric cancer. Cyclooxygenase-2 (COX-2) plays a crucial role in cancer progression. However, there is no experimental study on the regulation mechanism of EBV on COX-2 in EBVaGC. To understand more about the tumorigenic mechanism of EBVaGC, the study investigated the role of EBV encode latent membrane protein LMP1 and LMP2A in the regulation of COX-2. The expression of COX-2 was examined in EBVaGC and EBV negative gastric cancer (EBVnGC) cell lines. The plasmids were transfected in SGC7901 to overexpress LMP1/2A. Small interfering RNA (si-RNA) targeting LMP1/2A in GT38 and targeting TRAF2 in SGC7901 were used to detect the expression of COX-2. Furthermore, si-ERK1/2 and the MEK inhibitor PD0325901 were used to investigate whether p-ERK participate in the regulation of COX-2 in SGC7901. The overexpression of LMP1 or LMP2A in SGC7901 down-regulates both COX-2 and TRAF2 expression, and knockdown of LMP1 or LMP2A in GT38 resulted in a certain recovery of COX-2 and TRAF2 expression. Moreover, si-TRAF2 indicated that a sharp down-regulation of COX-2. And the decrease of p-ERK also mediates the inhibitory effect of LMP1 on COX-2. In summary, overexpression of LMP1 and LMP2A inhibits COX-2, which is mediated by a decrease of TRAF2, and p-ERK is involved in the inhibition of COX-2 by LMP1 in gastric cancer. [ABSTRACT FROM AUTHOR]