Single cell amplification methods for the study of cancer and cellular ageing.
Accumulation of somatic mutations takes place in ageing organisms and during malignant cell transformation and is accompanied by changes in cellular phenotype and function. The genomic damage that underlies the ageing phenotype is thought to occur stochastically. In cancer some changes causing cellular deregulation will be selected by clonal expansion but due to increased genetic instability it is still unknown which aberrations initiated sporadic cancer development. Thus, in ageing and cancer research detection of stochastically arising mutational events within a genome of 3x10(9) base pair requires global screening techniques and the analysis on a single cell level in contrast to pooled DNA or pooled RNA. We therefore focus on the development of techniques that enable the genome wide study of single cells. We successfully applied global amplification methods to study DNA and mRNA of single cancer cells that disseminated to the bone marrow of carcinoma patients early in disease progression. Our results emphasize the importance to study age-related diseases such as cancer on the level of single cells.
Klein
Christoph A
CA
Institut für Immunologie der Ludwigs-Maximilian Universität München, München 80336, Germany. christoph.klein@ifi.med.uni-muenchen.de
D000818:Animals; D018929:Cell Culture Techniques; D016922:Cellular Senescence / Q000235:genetics* / Q000502:physiology; D002869:Chromosome Aberrations; D016678:Genome; D006801:Humans; D009369:Neoplasms / Q000235:genetics* / Q000473:pathology / Q000503:physiopathology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000235:genetics*; D014407:Tumor Cells, Cultured
D012333:RNA, Messenger