Identification of pathways involved in aneuploidy onset and its tolerance using a DNA microarray approach

Global DNA Hypomethylation following 5-aza-2'-deoxycytidine treatment induces aneuploidy in HCT-116 tumor cells.

Aneuploidy, the alteration of the normal number of chromosomes, is found in most of the human solid tumors and correlated with to defects in the process of chromosome segregation (1). It was also suggested that the alteration of the 5-methylcytosine (5-mC) pattern in the chromosome pericentromeric region, generated to aneuploid cells (2, 3). To investigate the relationship between hypomethylation and whole chromosome aneuploidy, we treated HCT-116 cells, a near diploid line, with the demethylating agent 5-aza- 2'-deoxycytidine (DAC). The treatment with DAC for 24, 48 and 72 hours produced a progressive reduction of DNA methylation as shown by decrease of 5-mC signal. DNA hypomethylation res…

Identification of pathways involved in aneuploidy onset and its tolerance using a DNA microarray approach

Genomic instability is a hallmark of the majority of human tumors explaining the heterogeneity shown by tumor cells. This phenomenon is often associated with chromosomal instability (CIN) and aneuploidy, a condition in which tumor cells lose or gain chromosomes. Previously, we showed that posttranscriptional silencing by RNAi of pRb1, DNMT12 and MAD2 is associated with aneuploidy in cultured human cells reinforcing the idea that there are several roads leading to aneuploidy. In the attempt to understand if a common molecular signature exists underlying aneuploidy and its tolerance in tumor cells, we induced aneuploidy in human fibroblasts (IMR90) by depleting Rb, MAD2 and DNMT1 genes and an…

DNA MICROARRAY AND BIOINFORMATICS AS TOOLS TO IDENTIFY A COMMON MOLECULAR SIGNATURE SHARED BY HUMAN ANEUPLOID CELLS

Genomic instability is a hallmark of the majority of human tumors explaining the heterogeneity shown by tumor cells. This phenomenon is often associated with chromosomal instability (CIN) and aneuploidy, a condition in which tumor cells lose or gain chromosomes. Previously, we showed that posttranscriptional silencing by RNAi of pRb(1), DNMT1(2) and MAD2(3) is associated with aneuploidy in cultured human cells reinforcing the idea that there are several roads leading to aneuploidy. In the attempt to understand if a common molecular signature exists that underlies aneuploidy and its tolerance in tumor cells, we did post transcriptional silencing of Rb, MAD2 and DNMT1 in human fibroblasts (IM…

p14ARFPrevents Proliferation of Aneuploid Cells by Inducing p53-Dependent Apoptosis

Weakening the Spindle Assembly Checkpoint by reduced expression of its components induces chromosome instability and aneuploidy that are hallmarks of cancer cells. The tumor suppressor p14ARF is overexpressed in response to oncogenic stimuli to stabilize p53 halting cell progression. Previously, we found that lack or reduced expression of p14ARF is involved in the maintenance of aneuploid cells in primary human cells, suggesting that it could be part of a pathway controlling their proliferation. To investigate this aspect further, p14ARF was ectopically expressed in HCT116 cells after depletion of the Spindle Assembly Checkpoint MAD2 protein that was used as a trigger for aneuploidy. p14ARF…

Aneuploid IMR90 cells induced by depletion of pRB, DNMT1 and MAD2 show a common gene expression signature

Chromosome segregation defects lead to aneuploidy which is a major feature of solid tumors. How diploid cells face chromosome mis-segregation and how aneuploidy is tolerated in tumor cells are not completely defined yet. Thus, an important goal of cancer genetics is to identify gene networks that underlie aneuploidy and are involved in its tolerance. To this aim, we induced aneuploidy in IMR90 human primary cells by depleting pRB, DNMT1 and MAD2 and analyzed their gene expression profiles by microarray analysis. Bioinformatic analysis revealed a common gene expression profile of IMR90 cells that became aneuploid. Gene Set Enrichment Analysis (GSEA) also revealed gene-sets/pathways that are …