Precise breakpoint characterization of the colon adenocarcinoma cell line HT‐29 clone 19A by means of 24‐color fluorescence in situ hybridization and multicolor banding

Kuechler, Alma; Weise, Anja; Michel, Susanne; Schaeferhenrich, Anja; Pool‐Zobel, Beatrice L.; Claussen, Uwe; Liehr, Thomas

doi:10.1002/gcc.10163

Cited by 10 publications

(13 citation statements)

References 10 publications

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“…The following description of the HT29 cell line closely agrees with that of Kawai et al 28 and Kuechler et al 29 and differs from older reports. 30,31 An insertion of undefined material of chromosome 12 into the short arm of chromosome 3 was identified in our laboratory as well as an insertion of material of unknown origin into chromosome 9.…”

Section: Cell Lines Of the Cin Pathwaysupporting

confidence: 88%

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Genetic evolution in colon cancer KM12 cells and metastatic derivates

Camps

Morales

Prat

et al. 2004

Intl Journal of Cancer

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So far, CRC cell lines have contributed to descriptions of 2 patterns of genetic instability, affecting either microsatellite sequences or chromosome number and structure. Often, these patterns are mutually exclusive; while near-diploid karyotypes usually appear with MSI and chromosomal stability, near-triploid or tetraploid cells display a high degree of CIN and are stable at the microsatellite level. In the present study, we describe the genomic instability pattern of KM12 CRC cells. KM12C and derived cell lines with different metastatic properties were analyzed by conventional cytogenetics, CGH and M-FISH. Results were compared to 5 cell lines usually used as model of MSI and CIN. Concordance between our results and previously published SKY data are also reviewed. Interestingly, the poorly metastatic KM12C cell line displayed a near-diploid karyotype with high levels of structural chromosome instability and microsatellite instability. Most carcinomas show highly complex karyotypes, suggesting that one of the tumor-development events may be related to genomic instability. 1,2 Up to now, MSI has been the only type of genomic instability well characterized in a minority of tumors (about 15% of sporadic CRCs). These tumors show a deficiency in mismatch repair genes with a replication error phenotype and a stable near-diploid karyotype. 3-7 chromosome instability has been described in some CRC cell lines, resulting in numerical changes 8 (CIN) or structural rearrangements. 9 Often, these patterns are mutually exclusive; while near-diploid karyotypes usually appear with MSI and chromosome instability, near-triploid or tetraploid cells display a high degree of CIN and are stable at the microsatellite level.It has been postulated that primary alterations in solid tumors consist of submicroscopic mutations, whereas unbalanced chromosomal rearrangements, whether numerical or structural, are secondary events and most likely associated with selective pressures during tumor progression. 10 A previous report demonstrated clonal chromosomal aberrations in 87% of CRC tumors analyzed. 11 The most frequent numerical changes were, in order of decreasing frequency, ϩ7, -18, -14, ϩ13. Gain of chromosome 8q and loss of 8p, loss of 17p and gain of 17q as well as loss of the entire chromosome 22 are also unbalanced rearrangements that have been reported. 11 Based on cytogenetic alterations, Dutrillaux 12 proposed 3 different evolutionary pathways: monosomic-type tumors undergo structural rearrangements resulting in gains and losses of different chromosome arms and show a relatively high rate of endoreduplication, leading to the formation of hypotetraploid clones that evolve to near-triploid cells; trisomic cells are characterized by tumors with a progressive increase in chromosome number but accompanied by a scarce number of rearrangements; tumors that display a near-normal karyotype and MSI were grouped into the normal type. Unbalanced chromosomal abnormalities would be secondary events and most likely associated with selective pre...

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Section: Cell Lines Of the Cin Pathwaysupporting

confidence: 88%

“…51ϳ59͗3n͘,XX,ϪX [28],ϪY [28],Ϫ1 [28],t(1;9)(q12;q11) [27],der(2)t(2;12)(q37;q13) [27], del(3)(q12) [27],Ϫ4 [27],Ϫ5 [29],der(5)t(5;20)(q13;q13) [29],Ϫ6 [29],Ϫ7 [4],der(7)t(7;13)(q13;q?) [24], ϩder(7)t[inv (7);14][(q22q36)(q22;q22) [25],der(8)t(8;9)(q12;p13) [28],der(8)t(8;19)(q12;??)…”

Section: Cell Lines Of the Mis Pathwayunclassified

Genetic evolution in colon cancer KM12 cells and metastatic derivates

Camps

Morales

Prat

et al. 2004

Intl Journal of Cancer

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“…Recently, we demonstrated that a comprehensive analysis of complex karyotypes present in tumor cell lines can be done by a combination of M-FISH and chromosome-specific MCB (Kuechler et al, 2003). However, the strategy used in that report had the disadvantage, that -including the M-FISH study -up to 25 FISH experiments were necessary to come to a final result.…”

Section: Discussionmentioning

confidence: 99%

“…In previous studies the accuracy of the MCB technique in the determination of chromosomal breakpoints has been proven by comparison with M-FISH (Kuechler et al, 2003), CGH (Starke et al, 2001a;Tönnies et al, 2001;Stumm et al, 2002), microdissection plus reverse painting (Starke et al, 2001a, b) and locus and/or region-specific probes (Starke et al, 2001a;Liehr et al, 2002a;Weise et al, 2002). Two sets of chromosome specific MCB probes were assembled and used simultaneously before .…”

Section: Discussionmentioning

confidence: 99%

Multitude multicolor chromosome banding (mMCB) – a comprehensive one-step multicolor FISH banding method

Weise

Heller

Starke

et al. 2003

Cytogenet Genome Res

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Multicolor chromosome banding (MCB) using one single chromosome-specific MCB probe set per experiment was previously reported as powerful tool in molecular cytogenetics for the characterization of all kinds of human marker chromosomes. However, a quick analysis of karyotypes with highly complex chromosomal changes was hampered by the problem that up to 24 MCB experiments were necessary for a comprehensive karyotype description. To overcome that limitation the 138 available region-specific microdissection-derived libraries for all human chromosomes were combined to one single probe set, called multitude MCB (mMCB). A typical fluorescence banding pattern along the human karyotype is produced, which can be evaluated either by transforming these profiles into chromosome region-specific pseudo-colors or more reliably by studying the fluorescence profiles. The mMCB probe set has been applied on chromosomes of normal male and female probands, two primary myelodysplastic syndromes and two solid tumor cell lines. Additionally, a cell line of Gorilla gorilla (GGO) studied previously by single chromosome-specific MCB was reevaluated by the mMCB method. All results were in concordance with those obtained in parallel or by other cytogenetic and molecular cytogenetic approaches indicating that mMCB is a powerful multicolor FISH banding tool for fast characterization of complex karyotypes.

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“…Only studies with multicolour FISH and/or comparative genomic hybridisation descriptions of at least one of the fi ve cell lines were included for comparison (Masramon et al, 2000;Melcher et al, 2000Melcher et al, , 2002Abdel-Rahman et al, 2001;Tsushimi et al, 2001;Kawai et al, 2002;Kuechler et al, 2003;Camps et al, 2004;Kleivi et al, 2004;Mohr and Illmer, 2005).…”

Section: Data For Comparative Studiesmentioning

confidence: 99%

Cryptic terminal chromosome rearrangements in colorectal carcinoma cell lines detected by subtelomeric FISH analysis

Stewénius

Tanke

Wiegant

et al. 2006

Cytogenet Genome Res

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Epithelial tumour karyotypes are often difficult to study by standard cytogenetic methods because of poor chromosome preparation quality and the high complexity of their genomic rearrangements. Subtelomeric fluorescence in situ hybridisation (FISH) has proved to be a useful method for detecting cryptic constitutional chromosomal rearrangements but little is known about its usefulness for tumour cytogenetic analysis. Using a combination of chromosome banding, multicolour karyotyping and subtelomeric FISH, five colorectal cancer cell lines were characterised. The resulting data were compared to results from previous studies by comparative genomic hybridisation and spectral karyotyping or multicolour FISH. Subtelomeric FISH made it possible to resolve several highly complex chromosome rearrangements, many of which had not been detected or were incompletely characterised by the other methods. In particular, previously undetected terminal imbalances were found in the two cell lines not showing microsatellite instability.

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Precise breakpoint characterization of the colon adenocarcinoma cell line HT‐29 clone 19A by means of 24‐color fluorescence in situ hybridization and multicolor banding

Cited by 10 publications

References 10 publications

Genetic evolution in colon cancer KM12 cells and metastatic derivates

Genetic evolution in colon cancer KM12 cells and metastatic derivates

Multitude multicolor chromosome banding (mMCB) – a comprehensive one-step multicolor FISH banding method

Cryptic terminal chromosome rearrangements in colorectal carcinoma cell lines detected by subtelomeric FISH analysis

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