Combined Analysis of Murine and Human Microarrays and ChIP Analysis Reveals Genes Associated with the Ability of MYC To Maintain Tumorigenesis

Abstract

The MYC oncogene has been implicated in the regulation of up to thousands of genes involved in many cellular programs including proliferation, growth, differentiation, self-renewal, and apoptosis. MYC is thought to induce cancer through an exaggerated effect on these physiologic programs. Which of these genes are responsible for the ability of MYC to initiate and/or maintain tumorigenesis is not clear. Previously, we have shown that upon brief MYC inactivation, some tumors undergo sustained regression. Here we demonstrate that upon MYC inactivation there are global permanent changes in gene expression detected by microarray analysis. By applying StepMiner analysis, we identified genes whose expression most strongly correlated with the ability of MYC to induce a neoplastic state. Notably, genes were identified that exhibited permanent changes in mRNA expression upon MYC inactivation. Importantly, permanent changes in gene expression could be shown by chromatin immunoprecipitation (ChIP) to be associated with permanent changes in the ability of MYC to bind to the promoter regions. Our list of candidate genes associated with tumor maintenance was further refined by comparing our analysis with other published results to generate a gene signature associated with MYC-induced tumorigenesis in mice. To validate the role of gene signatures associated with MYC in human tumorigenesis, we examined the expression of human homologs in 273 published human lymphoma microarray datasets in Affymetrix U133A format. One large functional group of these genes included the ribosomal structural proteins. In addition, we identified a group of genes involved in a diverse array of cellular functions including: BZW2, H2AFY, SFRS3, NAP1L1, NOLA2, UBE2D2, CCNG1, LIFR, FABP3, and EDG1. Hence, through our analysis of gene expression in murine tumor models and human lymphomas, we have identified a novel gene signature correlated with the ability of MYC to maintain tumorigenesis.

Author Summary

The targeted inactivation of oncogenes may be a specific and effective treatment of cancer. However, how oncogene inactivation leads to tumor regression is not clear. Previously, we have shown that even the brief inactivation of the MYC oncogene can result in the sustained regression of at least some tumors. To understand the mechanism, we have utilized several novel genomic analyses to define a set of genes that strongly correlate with the ability of the MYC oncogene to maintain tumorigenesis. First, we generated a novel data set from microarray analyses of murine tumors that we analyzed by StepMiner to identify discrete step changes in gene expression after the inactivation or the reactivation of the MYC oncogene. Second, we utilized Boolean Network Analysis to further define the subset of genes highly correlated with MYC in human tumorigenesis. Third, we utilized ChIP analysis to demonstrate that in many cases the permanent changes of gene expression we uncovered were associated with changes in the ability of MYC to occupy the promoter locus. Our general strategy could be similarly utilized in other experimental model systems to understand how specific oncogenes contribute to the maintenance of tumorigenesis.