Amplification of the pericentromeric region of chromosome 1 in a newly established colon carcinoma cell line.

The LRWZ cell line was established from an ascitic effusion of a colon adenocarcinoma. We studied the karyotype of LRWZ cells using G-banding and chromosome painting. The cell line is near triploid and is characterized by several chromosome rearrangements and pronounced intermetaphase variation. Chromosome painting probes revealed numerous labeled regions on different chromosomes, indicating that several translocations occurred during the evolution of the cell population. The 10 recurrent marker chromosomes identified (M1-M10) were derived from complex rearrangements involving up to three different chromosomes. M2 is a particularly interesting marker that originated from the amplification of the pericentromeric region of chromosome 1 and has a peculiar organization comprising five copies of the region included between 1p21 and 1q21 and is surprisingly stable: it is present in all the metaphases analyzed, has telomeric DNA at both termini, and contains one active and four inactivated centromeres. To provide insights into the molecular mechanisms that generated M2, we performed fluorescence in situ hybridization experiments using a panel of probes mapping near the centromere of chromosome 1 and three probes for different satellite sequences; the formation of chromosome M2 required the intervention of several rearrangements including unequal exchange, chromatid breakage followed by fusion of the sister chromatids, and loss of centromeric heterochromatin.