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Epstein-Barr Virus (EBV) Latent Membrane Protein 1 (LMP1) Regulates Host B Cell miR-155 and its Target FOXO3a via PI3K p110a Activation

ABSTRACT: The purpose of this study was to investigate the host cell miRs regulated by LMP1 from the B95-8 strain of EBV. In this study, we utilized an established chimeric, inducible NGFRLMP1 siganling molecule stably expressed in EBV- BL41 Burkitt’s Lymphoma cell lines (BL41 NGFRLMP1 cells). A qPCR array was used to measure the expression level of 377 previously identified miRs from RNA isolated from two sources: BL41 NGFRLMP1 cells in which LMP1 signaling was not activated (n=2, control) and BL41 NGFRLMP1 cells in which LMP1 signaling was activated for 12 hours (n=2).

ORGANISM(S): Homo sapiens

PROVIDER: GSE136721 | GEO | 2019/10/28

REPOSITORIES: GEO

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Project description:The Epstein Barr virus (EBV) encoded latent membrane protein-1 (LMP1) is a functional homologue of the tumor necrosis factor receptor family and contributes substantially to the oncogenic potential of EBV through activation of Nuclear Factor-kappaB (NF-kappaB). MicroRNAs (miRNAs) are a class of small RNA molecules that are involved in the regulation of cellular processes such as growth, development, and apoptosis, and have recently been linked to cancer phenotypes. Through miRNA microarray analysis, we demonstrate that LMP1 dysregulates the expression of several cellular miRNAs, including the most highly regulated of these, miR-146a. Quantitative RT-PCR analysis confirmed induced expression of miR-146a by LMP1. Analysis of miR-146a expression in EBV latency type III and type I cell lines revealed substantial expression of miR-146a in type III (which express LMP1) but not in type I cell lines. Reporter studies demonstrated that LMP1 induces miR-146a predominantly through two NF-kappaB binding sites in the miR-146a promoter and identified a role for an OCT-1 site in conferring basal and induced expression. Array analysis of cellular mRNAs expressed in Akata cells transduced with an miR-146a expressing retrovirus identified genes that are directly or indirectly regulated by miR-146a, including a group of interferon responsive genes that are inhibited by miR-146a. Since miR-146a is known to be induced by agents that activate the interferon response pathway (including LMP1), these results suggest that miR-146a functions in a negative feedback loop to modulate the intensity and/or duration of the interferon response. Keywords: microRNA expression modified by EBV encoded oncogene, LMP1 EBV negative Mutu E1dn Cl. 3 cells were transduced with either a control retrovirus (pEhyg) or an LMP1 containing retrovirus (pEhyg-FLAG-LMP1). Cells were selected with hygromycin for approximately 2 weeks afterwhich 5 pairs of RNA preps were carried out on 5 successive days. Arrays GSM255656 and GSM255659 are dye swaps in which LMP1 samples were labeled with Cy3 and control samples were labeled with Cy5.

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Epstein-Barr Virus (EBV) Latent Membrane Protein 1 (LMP1) Regulates Host B Cell miR-155 and its Target FOXO3a via PI3K p110a Activation

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