Impact of persistent cytomegalovirus infection on human neuroblastoma cell gene expression

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Abstract

In a model of human neuroblastoma (NB) cell lines persistently infected with human cytomegalovirus (HCMV) we previously showed that persistent HCMV infection is associated with an increased malignant phenotype, enhanced drug resistance, and invasive properties. To gain insights into the mechanisms of increased malignancy we analyzed the global changes in cellular gene expression induced by persistent HCMV infection of human neuroblastoma cells by use of high-density oligonucleotide microarrays (HG-U133A, Affymetrix) and RT-PCR. Comparing the gene expression of different NB cell lines with persistently infected cell sub-lines revealed 11 host cell genes regulated in a similar manner throughout all infected samples. Nine of these 11 genes may contribute to the previously observed changes in malignant phenotype of persistently HCMV infected NB cells by influencing invasive growth, apoptosis, angiogenesis, and proliferation. Thus, this work provides the basis for further functional studies.

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Materials and methods

Cell cultures and virus strains. Human NB cell line MHH-NB-11 was obtained by the “Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH” (DSMZ, Braunschweig, Germany). The human NB cell lines UKF-NB-2, UKF-NB-3, and UKF-NB-4 were established as described before [12]. SH-SY5Y was obtained from ATTC/LGC (Promochem, Wesel, Germany). All cells were propagated in Iscove’s modified Dulbecco’s medium (IMDM, Seromed, Berlin, Germany) supplemented with 10% fetal calf serum (FCS, Gibco, Karlsruhe,

Alteration in gene expression by productive persistent infection of NB cells

A microarray approach was applied to assess changes in host cell gene expression in response to persistent HCMV infection. Focusing on the general impact of persistent HCMV infection on neuroblastoma cell lines we compared the gene expression of several parental NB cell lines (MHH-NB-11, UKF-NB-4, and UKF-NB-2) to their persistently AD169 or Hi91-infected sub-cell lines (MHH-NB-11AD169, UKF-NB-2AD169, UKF-NB-4AD169, MHH-NB-11Hi91, and UKF-NB-4Hi91). Applying strict filter algorithms followed by

Acknowledgments

This work was supported by the friendly organization “Hilfe für krebskranke Kinder Frankfurt e.V.” and its foundation “Frankfurter Stiftung für krebskranke Kinder.” The authors wish to express their sincere thanks Rouslan Kotchetkov for fruitful discussion, to Lena Stegmann and Gesa Meincke for their skillful technical assistance.

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