XMRV Infection Induces Host Genes that Regulate Inflammation & Cellular Physiology

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http://www.jurology.com/article/S0022-5347%2811%2900620-3/fulltext#sec0005

The Journal of Urology

Volume 185, Issue 4, Supplement, Page e113,

April 2011

XMRV INFECTION INDUCES HOST GENES

THAT REGULATE INFLAMMATION AND

CELLULAR PHYSIOLOGY

Lee*, Elona Gusho, Jaydip Das

Gupta, Klein, Silverman,

Cleveland, OH

INTRODUCTION AND OBJECTIVES

XMRV is a novel human retrovirus associated with

prostate cancer. Although other gammaretroviruses

cause cancer in animals, it remains unknown if XMRV

is a cause of disease.

However, indirect or direct modes of carcinogenesis

by XMRV have been suggested depending on whether

the virus was found in stroma or malignant

epithelium.

To gain insight into the possible role of XMRV in

these diseases we have identified genes that are

induced in response to XMRV infection.

METHODS

Prostate cancer cell line DU145 was infected for 8,

24, 48 and 120h with XMRV. A comparison to

uninfected DU145 cells cultured for the same periods

of time served as controls.

A population of total RNA was isolated using Qiagen

RNeasy Mini Kit followed by digestion of DNA with

DNAse treatment.

XMRV infections at the different time points were

monitored using real-time RT-PCR for env XMRV RNA.

The RNA samples were analyzed for gene expression

using Sentrix humanRef-8 v3 expression bead chips

from Illumina (Cleveland Clinic Genomics Core).

To verify the results obtained by the array

experiment, we determined induction of a subset of

the regulated genes.

Total RNA was reverse transcribed to cDNA using

iScript Select cDNA Synthesis Kit from Bio-Rad

(random primers method).

Induction of selected genes by XMRV infection was

verified by qPCR (Relative Quantification) from the

cDNA pool using SYBR Green master mix.

Fold-induction at each time point for the individual

mRNAs was determined.

In addition, pathway predictions were determined

using Ingenuity Systems (content version 3002)

software for genes induced by more than 2-fold

following XMRV infection.

RESULTS

In gene expression profiling, we observed maximal

gene induction between 24 and 48 h post-infection.

For example, the pro-inflammatory cytokine IL8

gene, a potential contributing factor to androgen

independent growth of late-stage prostate cancer,

was consistently induced by XMRV infection by up to

6-fold.

Of the XMRV induced genes, pathway analysis

indicated 10 genes are implicated in cell morphology,

11 genes in cellular development, 12 genes in

cell-to-cell signaling and interaction, 11 genes in

cellular movement and 13 genes in cellular growth

and proliferation.

CONCLUSIONS

The chemokine IL-8 is one of the most highly

induced genes in response to XMRV infection of

prostate cancer cell line DU145.

XMRV induction of the 30 host genes identified in

this study suggests a profound effect of the virus on

fundamental cellular physiology and inflammation.

These findings could be relevant to the possible

pathogenic effects on XMRV in prostate cancer.

Source of Funding: None

Copyright © 2011 Elsevier, Inc. All rights reserved