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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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HMF3A_RNAi

ABSTRACT: HMF3A cells, created from adult human mammary fibroblasts by immortalisation with a thermolabile SV40 large T antigen and the catalytic sub-unit of human telomerase, undergo co-ordinated induction of cellular senescence upon inactivation of T antigen. The pSUPER-retro vector system (Brummelkamp, 2002) was used to selectively target genes for mRNA degradation in an attempt to determine if they had a role, in the changes to the transcriptome upon LT inactivation. The genes chosen for targeting were BTG2, NR4A3, DUSP1, PHLDA1 and STACb. Keywords = LT antigen Keywords = senescence Keywords = fibroblast Keywords = BTG2 Keywords = DUSP1 Keywords = MKP-1 Keywords = NR4A3 Keywords = STAC Keywords = PHLDA1 Keywords = RNAi

ORGANISM(S): Homo sapiens

SUBMITTER: Parmjit Jat

PROVIDER: E-GEOD-690 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Transcriptional networks and cellular senescence in human mammary fibroblasts.

Hardy K K Mansfield L L Mackay A A Benvenuti S S Ismail S S Arora P P O'Hare M J MJ Jat P S PS

Molecular biology of the cell 20041201 2

Senescence, the molecular program that limits the finite proliferative potential of a cell, acts as an important barrier to protect the body from cancer. Techniques for measuring transcriptome changes and for modulating their expression suggest that it may be possible to dissect the transcriptional networks underlying complex cellular processes. HMF3A cells are conditionally immortalized human mammary fibroblasts that can be induced to undergo coordinated senescence. Here, we used these cells in ...[more]

PMID: 15574883

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Project description:HMF3A cells, created from adult human mammary fibroblasts by immortalisation with a thermolabile SV40 large T antigen and the catalytic sub-unit of human telomerase, undergo co-ordinated induction of cellular senescence upon inactivation of T antigen. HMF3A cells cease proliferating by 4 days at 39Â°C. We directly compared the gene expression profiles of cells at 33.5Â°C and after shift up to 39.5Â°C for 7 days (10 arrays, 3 biological replicates). To eliminate genes that change in response to the temperature shift, we compared the profiles of HMF3Dwt cells at 33Â°C and those shifted up to 39Â°C for 7 days (4 arrays, 2 biological replicates). Since HMF3Dwt cells were immortalised with wild type U19 LT antigen, they proliferate at both temperatures. To break up the list of genes obtained with the 7 day shift, we performed several further comparisons on the arrays. HMF3A cells at 33Â°C were compared to the primary donor fibroblasts (HMF3 passage 8, 4 arrays), HMF3A cells shifted to 39Â°C for 2 days (4 arrays, 2 biological repeats), HMF3A cells shifted to 39Â°C for 7 days and then shifted back to 33Â°C for 3 days (8 arrays, 4 biological repeats) or 7 days (4 arrays, 2 biological repeats). As a comparison to the irreversible process of senescence, we sought to identify genes that changed upon reversible growth arrest, quiescence. Thus changes that occurred when the cells were grown to confluency and starved were also determined (2 biological replicate, 6 arrays). Keywords = Rb Keywords = fibroblast Keywords = immortalization Keywords = LT antigen Keywords = p53 Keywords = quiescence Keywords = senescence

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