Expression Profile of MicroRNAs in Epstein-Barr Virus-Infected AGS Gastric Carcinoma Cells

Marquitz, Aron R.; Mathur, Anuja; Chugh, Pauline; Dittmer, Dirk P.; Raab‐Traub, Nancy

doi:10.1128/jvi.02662-13

Cited by 84 publications

(91 citation statements)

References 18 publications

(19 reference statements)

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“…1C). The changes in expression observed predicted that the infected cells would have an increase in proliferation and cell viability and a decrease in apoptosis and cell death, which is consistent with the increased anchorage-independent growth previously observed in these cell lines (4,15). In order to determine whether the transcription changes were caused by a discrete subset of activators and/or repressors, the Ingenuity Pathway Analysis (IPA) software package was used to query possible upstream regulators that could account for these transcription changes.…”

Section: Resultssupporting

confidence: 84%

“…In order to better understand the effects of latent EBV infection in epithelial cells, RNA-seq was used to obtain a complete transcriptional profile of both uninfected and infected AGS cells. mRNA libraries were generated using the Illumina TruSeq RNA prep kit to obtain libraries from RNA isolated from three preparations of uninfected AGS cells, one EBV-infected cell line obtained that has been previously profiled by microarray (4), and two additional EBV-infected AGS cell lines that have been recently generated (15). Sequence reads generated by paired-end, 100-bp sequencing on the Illumina HiSeq 2000 instrument were aligned to both the human (hg19) and the Akata EBV (KC207813) genomes by using the splicing aware read aligner Tophat.…”

Section: Resultsmentioning

confidence: 99%

“…AGS cells infected with EBV express high levels of the BART miRNAs (15), and previous profiling of one AGS line by microarray suggested that a substantial subset of the downregulated genes after latent infection were direct targets of the BART miRNAs (4). To confirm that the BART miRNAs contribute to the changes in gene expression profiled in the multiple infected cell lines in this report, the genes changed in expression were overlapped with the most extensive lists of BART miRNA targets published to date, where the BART miRNA targets were identified by PAR-CLIP in a peripheral effusion lymphoma cell line dually infected with EBV and KSHV, as well as the NPC cell line C666 (25,26).…”

Section: Resultsmentioning

confidence: 99%

“…The gastric carcinoma cell line AGS was grown in F-12 media (Life Technologies, Inc.) with 10% fetal bovine serum and antibiotic/antimycotic (Life Technologies, Inc.). The generation of AGS cells clonally infected with EBV Akata BX1 was previously described (15). AGS-EBV cells were also maintained with 500 g of G418 (Life Technologies, Inc.)/ml to select for retention of the EBV episome.…”

Section: Methodsmentioning

confidence: 99%

“…Importantly, EBV infection of AGS cells consistently induces anchorage-independent growth and increased motility (4,15,16). Gene expression microarray analysis of a single AGS-EBV cell line compared to parental cells suggested that there are widespread changes in host transcription due to latent EBV infection and that the downregulated genes were highly enriched for BART miRNA targets (4).…”

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confidence: 99%

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Host Gene Expression Is Regulated by Two Types of Noncoding RNAs Transcribed from the Epstein-Barr Virus BamHI A Rightward Transcript Region

Marquitz

Mathur

Edwards

et al. 2015

J Virol

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In Epstein-Barr virus-infected epithelial cancers, the alternatively spliced BamHI A rightward transcripts (BARTs) are the most abundant viral polyadenylated RNA. The BART introns form the template for the production of 44 microRNAs (miRNAs), and the spliced and polyadenylated exons form nuclear non-protein-coding RNAs. Analysis of host cell transcription by RNA-seq during latency in AGS cells identified a large number of reproducibly changed genes. Genes that were downregulated were enriched for BART miRNA targets. Bioinformatics analysis predicted activation of the myc pathway and downregulation of XBP1 as likely mediators of the host transcriptional changes. Effects on XBP1 activity were not detected in these cells; however, myc activation was confirmed through use of a myc-responsive luciferase reporter. To identify potential regulatory properties of the spliced, polyadenylated BART RNAs, a full-length cDNA clone of one of the BART isoforms was obtained and expressed in the Epstein-Barr virus (EBV)-negative AGS cells. The BART cDNA transcript remained primarily nuclear yet induced considerable and consistent changes in cellular transcription, as profiled by RNA-seq. These transcriptional changes significantly overlapped the transcriptional changes induced during latent EBV infection of these same cells, where the BARTs are exclusively nuclear and do not encode proteins. These data suggest that the nuclear BART RNAs are functional long noncoding RNAs (lncRNAs). The abundant expression of multiple forms of noncoding RNAs that contribute to growth regulation without expression of immunogenic proteins would be an important mechanism for viral oncogenesis in the presence of a functional immune system. IMPORTANCEInfection with Epstein-Barr virus (EBV) is nearly ubiquitous in the human population; however, it does contribute to the formation of multiple types of cancer. In immunocompromised patients, EBV causes multiple types of lymphomas by expressing viral oncogenes that promote growth and survival of infected B lymphocytes. EBV-positive gastric carcinoma does not require immune suppression, and the viral oncoproteins that are frequent targets for an immunological response are not expressed. This study demonstrates using transcriptional analysis that the expression of various classes of viral non-protein-coding RNAs likely contribute to the considerable changes in the host transcriptional profile in the AGS gastric cancer cell line. This is the first report to show that the highly expressed polyadenylated BamHI A rightward transcripts (BART) viral transcript in gastric carcinoma is in fact a functional viral long noncoding RNA. These studies provide new insight into how EBV can promote transformation in the absence of viral protein expression. L atent infection with Epstein-Barr virus (EBV), a nearly ubiquitous human herpesvirus, is responsible for multiple malignancies in the two types of cells that it infects: B lymphocytes and epithelial cells (1). The mechanism by which EBV leads to transformation dep...

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Section: Resultssupporting

confidence: 84%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Host Gene Expression Is Regulated by Two Types of Noncoding RNAs Transcribed from the Epstein-Barr Virus BamHI A Rightward Transcript Region

Marquitz

Mathur

Edwards

et al. 2015

J Virol

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Epstein‐Barr virus miR‐BART2‐5p and miR‐BART11‐5p regulate cell proliferation, apoptosis, and migration by targeting RB and p21 in gastric carcinoma

Zhao

Zhang

et al. 2022

Journal of Medical Virology

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Epstein‐Barr virus (EBV) was the first tumor virus discovered in humans and can cause various types of tumors. Molecular classification suggests that EBV‐associated gastric cancer (EBVaGC) is a unique subtype of gastric cancer.EBV was also the first virus found to encode its own microRNAs. However, the functions of many miRNAs remain unknown. This study investigated the roles and targets of miR‐BART2‐5p (BART2‐5p) and miR‐BART11‐5p (BART11‐5p) in EBVaGC. The expression of RB and p21 in EBVaGC and EBV negative GC (EBVnGC) cells was evaluated by western blotting. Expression of BART2‐5p and BART11‐5p in EBVaGC cells was evaluated by droplet digital PCR. The effects of BART2‐5p or BART11‐5p and their potential mechanisms were further investigated using cell counting kit‐8, colony formation assay, flow cytometry analysis, and transwell assay. BART2‐5p and BART11‐5p were abundantly expressed and RB and p21 were downregulated in EBVaGC cells. BART2‐5p regulates RB and p21 expression by directly targeting them. BART11‐5p regulates RB expression by directly targeting RB. Both BART2‐5p and BART11‐5p promoted proliferation and migration of gastric cancer cells, while inhibiting apoptosis and promoting S‐phase arrest of the cell cycle. Thus, BART2‐5p and BART11‐5p play important roles in promoting proliferation and migration, and inhibiting apoptosis in EBVaGC by targeting RB and p21, thus providing new potential therapeutic targets for EBVaGC.

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The role of Epstein–Barr virus in epithelial malignancies

Tsao

Tsang

et al. 2014

The Journal of Pathology

275

274

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The close association of Epstein–Barr virus (EBV) infection with non-keratinizing nasopharyngeal carcinomas and a subset of gastric carcinomas suggests that EBV infection is a crucial event in these cancers. The difficulties encountered in infecting and transforming primary epithelial cells in experimental systems suggest that the role of EBV in epithelial malignancies is complex and multifactorial in nature. Genetic alterations in the premalignant epithelium may support the establishment of latent EBV infection, which is believed to be an initiation event. Oncogenic properties have been reported in multiple EBV latent genes. The BamH1 A rightwards transcripts (BARTs) and the BART-encoded microRNAs (miR-BARTs) are highly expressed in EBV-associated epithelial malignancies and may induce malignant transformation. However, enhanced proliferation may not be the crucial function of EBV infection in epithelial malignancies, at least in the early stages of cancer development. EBV-encoded gene products may confer anti-apoptotic properties and promote the survival of infected premalignant epithelial cells harbouring genetic alterations. Multiple EBV-encoded microRNAs have been reported to have immune evasion functions. Genetic alterations in host cells, as well as inflammatory stroma, could modulate the expression of EBV genes and alter the growth properties of infected premalignant epithelial cells, encouraging their selection during carcinogenesis.

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Expression Profile of MicroRNAs in Epstein-Barr Virus-Infected AGS Gastric Carcinoma Cells

Cited by 84 publications

References 18 publications

Host Gene Expression Is Regulated by Two Types of Noncoding RNAs Transcribed from the Epstein-Barr Virus BamHI A Rightward Transcript Region

Host Gene Expression Is Regulated by Two Types of Noncoding RNAs Transcribed from the Epstein-Barr Virus BamHI A Rightward Transcript Region

Epstein‐Barr virus miR‐BART2‐5p and miR‐BART11‐5p regulate cell proliferation, apoptosis, and migration by targeting RB and p21 in gastric carcinoma

The role of Epstein–Barr virus in epithelial malignancies

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