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First published online 21 March 2006
doi: 10.1242/jcs.02871

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Genetic instability and divergence of clonal populations in colon cancer cells in vitro

¹ IDIBELL-Institut de Recerca Oncològica, L'Hospitalet, 08907 Barcelona, Spain
² IDIBELL-Institut Català d'Oncologia, L'Hospitalet, 08907 Barcelona, Spain
³ Departament de Biologia Cellular, Fisiologia i Immunologia, Institut de Biotecnologia i Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193 Barcelona, Spain

View larger version (17K): [in a new window]   Fig. 1. Phylogenetic tree of metaphases analyzed in parental and derived clones of SW480 cells. Each dot represents a cell. Parental cells are represented in red and show the smaller genetic distances. Cells of clones S1, S2 and S4 are represented in yellow, blue and green, respectively. The scale of the diversity index (Table 1) is shown at the bottom right.

View larger version (64K): [in a new window]   Fig. 2. Illustrative examples of the DNA fingerprinting analysis of the three cell lines with six different primers. Each panel comprises the parental cells (left-most lane) and different clones and subclones. The six different primers used are given above each panel (described further in Materials and Methods). Differences between parental and derived cells were analyzed densitometrically (see Materials and Methods).

View larger version (20K): [in a new window]   Fig. 3. Split decomposition tree (upper panel) and principal component analysis (PCA) (lower panel) constructed using AP-PCR data of clones and subclones compared with the respective parental cell. Parental cell lines were represented as a single and common point (marked P) defined by a vector (0,0,0,...). Each AP-PCR band corresponds to a component of the vector and the values indicate loss (–1), gain (+1), or no change (0) in the clone in regard to the fingerprint of the parental cell. Clones of the SW480 cell line are labeled `S', clones of LoVo are labeled `L' and clones of HCT116 are labeled `H'. Tree and space arrangement revealed the differential nature of genetic alterations characterizing the divergence in the clones derived from each cell line. In PCA analysis, the spikes are traced to the centroid point of the clones for each cell line.

View larger version (22K): [in a new window]   Fig. 4. Genetic divergence of clonal populations was of the same order in two successive cloning processes. The index of genetic divergence was assessed by AP-PCR (see Materials and Methods). The X-axis represents the divergence between each expanded cell population cloned directly from the parental cells (Cloning 1) versus (vs) the corresponding parental cells. The Y-axis represents the divergence between each of the second-generation clones (Cloning 2, expanded from a clone from the first cloning) versus the clone from which they were isolated. The diagonal represents ratio 1, corresponding to equal levels of genetic divergence in both cloning processes.