Analysis of Marker Chromosomes in Renal Cell Carcinoma Cell Line Caki-1 by Chromosome Microdissection

Shin, Jae Seop; Park, Soo Yeun; Chun, Yong Hyuck; Park, Sun Hwa

Abstract

PURPOSE: The analysis of recurring chromosome aberrations has become an integral part of the diagnostic and prognostic workup of many human cancers, and their molecular analyses have facilitated the identification of genes related to the pathogenesis of cancer But the technical limitation of conventional cytogenetic method makes unable us to characterize all recognizable chromosome rearrangements. The generation of chromosome region specific painting probe by PCR amplification of microdissected chromosomal DNA has proven extremely useful in identification of chromosomal derivation for marker chromosomes which are indeterminable by routine chromosome banding analysis. In this study we have constructed and analyzed the band-specific painting probe for unidentified marker chromosomes of renal cell carcinoma cell line, Caki-1 to determine the derivative chromosomes and the painting probes applied to CURC-II to compare the marker chromosomes.
MATERIALS AND METHODS
Microdissection was performed on 9q+ and unidentified one of Caki-1, and chromosomal BNAs were amplified by PCR using topoisomerase I and T7 DNA polymerase. Fluorescent in situ hybridization was conducted with biotin labeled PCR products to normal, Caki-1 and CURC- II metaphase chromosomes.
RESULTS
With this method, it was possible to construct the band-specific painting probes for markers and the probes hybridized specifically to the dissected regions and derivative chromosomes. The 9q+ and unidentified one were identified as t(9;17)(q34;q21) and t(15;20) respectively. The marker chromosomes - t(9;I 7), der(1 ;17)(ql0;q10), t(15;20), t(?;15), der(1 ;20), t(14;89) were examined same in Caki-1 and CURC-II.
CONCLUSIONS
Thus this methodological advance significantly extends the limits of conventional cytogenetic analysis by enabling the analysis of unknown chromosome regions, and these painting probes can be applied to detect the similar marker chromosomes in renal cell carcinoma, and the probe pools for markeys may be used to identify the cancer-relevant genes.