IPM-FISH, a new M-FISH approach using IRS-PCR painting probes: application to the analysis of seven human prostate cell lines.
We have developed an alternative multicolor karyotyping technique based on multiplex fluorescence in situ hybridization (M-FISH) and our own optical device with a specific filter set. The most innovative part of our development is the use of interspersed polymerase chain reaction (IRS-PCR) painting probes that show an R-band pattern simultaneous to the combinatorial labeling. This allows us not only to recognize the origin of chromosomal fragments, but to identify the breakpoints as well. We have used this technique to analyze seven cell lines: four prostate cancer cell lines (CA-HPV-10, LNCaP, DU145, and PC3), and three normal transformed epithelial prostate cell lines (PNT1B, PNT2, and PZ-HPV-7). In order to validate our IRS-PCR multiplex FISH (IPM-FISH) technique and to complement the results, we applied comparative genomic hybridization (CGH) and FISH analysis, showing good correlation with the IPM-FISH results. To date, molecular and cytogenetic studies have identified several chromosomal regions that are altered in human prostate cancer; several candidate genes have been suggested. However, reliable markers for predicting the aggressiveness of early prostate cancer are not yet available. Our results show several common, unbalanced rearrangements in the cell lines. These rearrangements are similar to regions already implicated in prostate cancer, validating these cell lines as a good model system.
Aurich-Costa
J
J
Cytogenetics Department, Genomic Research Center, GENSET, S.A., Evry, France. joan.aurich@gense.fr
Vannier
A
A
Grégoire
E
E
Nowak
F
F
Cherif
D
D
D002461:Cell Line, Transformed; D002871:Chromosome Banding; D020223:Chromosome Painting / Q000295:instrumentation / Q000379:methods; D015342:DNA Probes / Q000235:genetics*; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation / Q000379:methods*; D007621:Karyotyping; D008297:Male; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D011467:Prostate / Q000737:chemistry* / Q000166:cytology / Q000473:pathology; D011471:Prostatic Neoplasms / Q000737:chemistry / Q000235:genetics / Q000473:pathology; D015203:Reproducibility of Results; D014407:Tumor Cells, Cultured
D015342:DNA Probes; D005456:Fluorescent Dyes