Ankyrin G overexpression in Hutchinson-Gilford progeria syndrome fibroblasts identified through biological filtering of expression profiles.
Hutchinson-Gilford progeria syndrome (HGPS; MIM 176670) is a rare disease characterized by accelerated aging. In this study, light and immunofluorescence microscopy were used to assess morphological changes, measures of cell growth kinetics and gene expression profiles in HGPS cells and normal fibroblasts in culture. A filtering strategy was developed based on differentially expressed transcripts seen consistently across three culture stages based on cell passage number. This filtering strategy produced a list of 66 unique differentially expressed genes, of which approximately 40% were upregulated in HGPS cells compared to normal fibroblasts. The increased mRNA expression in HGPS cells that was seen for one gene defined using this strategy--namely ANK3--was validated using quantitative reverse-transcriptase amplification, Western analysis and immunofluorescence microscopy, all of which showed significantly increased ankyrin G expression. These findings demonstrate differences in morphology, growth kinetics and mRNA expression profiles in HGPS cells compared to normal fibroblasts in culture, including increased expression of ANK3/ankyrin G. Furthermore, other genes that co-clustered with ANK3 might provide mechanistic clues regarding senescence in cultured HGPS cells.
Wang
Jian
J
Vascular Biology Research Group, Robarts Research Institute, 100 Perth Drive, London, ON, Canada, N6A 5K8.
Robinson
John F
JF
Vascular Biology Research Group, Robarts Research Institute, 100 Perth Drive, London, ON, Canada, N6A 5K8.
O'Neil
Caroline H
CH
Vascular Biology Research Group, Robarts Research Institute, 100 Perth Drive, London, ON, Canada, N6A 5K8.
Edwards
Jane Y
JY
Vascular Biology Research Group, Robarts Research Institute, 100 Perth Drive, London, ON, Canada, N6A 5K8.
Williams
Christina M
CM
Vascular Biology Research Group, Robarts Research Institute, 100 Perth Drive, London, ON, Canada, N6A 5K8.
Huff
Murray W
MW
Vascular Biology Research Group, Robarts Research Institute, 100 Perth Drive, London, ON, Canada, N6A 5K8.|Department of Medicine, University of Western Ontario, London, ON, Canada.
Pickering
J Geoffrey
JG
Vascular Biology Research Group, Robarts Research Institute, 100 Perth Drive, London, ON, Canada, N6A 5K8.|Department of Medicine, University of Western Ontario, London, ON, Canada.
Hegele
Robert A
RA
Vascular Biology Research Group, Robarts Research Institute, 406-100 Perth Drive, London, ON, Canada, N6A 5K8. hegele@robarts.ca.|Department of Medicine, University of Western Ontario, London, ON, Canada. hegele@robarts.ca.
D017487:Ankyrins / Q000096:biosynthesis* / Q000235:genetics*; D002460:Cell Line; D049109:Cell Proliferation; D016000:Cluster Analysis; D005347:Fibroblasts / Q000378:metabolism*; D005786:Gene Expression Regulation; D016678:Genome; D006801:Humans; D007700:Kinetics; D008856:Microscopy, Fluorescence; D020411:Oligonucleotide Array Sequence Analysis; D011371:Progeria / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
C490613:ANK3 protein, human; D017487:Ankyrins; D012333:RNA, Messenger