Identification of chromosomal aberration in esophageal cancer cells by mixed BAC DNA probes of chromosome arms and regions
Received date: 2014-01-27
Revised date: 2014-03-14
Online published: 2014-05-28
Chromosomal aberration is an important genetic feature of malignant tumor cells. This study aimed to clarify whether BAC DNA could be used to identify chromosome region and arm alterations. For each chromosome region, five to ten 1 Mb BAC DNA clones were selected to construct mixed BAC DNA clones for the particular region. All of the mixed clones from regions which could cover the whole chromosome arm were then mixed to construct mixed BAC DNA clones for the arms. Mixed BAC DNA probes of arms and regions were labeled by degenerate oligonucleotide primed PCR (DOP-PCR) and Nick translation techniques, respectively. The specificities of these probes were validated by fluorescence in situ hybridization (FISH) on the metaphase chromosomes of normal human peripheral blood lymphocytes. FISH with arm-specific mixed BAC DNA probes showed that chromosomal rearrangements and involved chromosome arms were confirmed in several esophageal cancer cells. By using region-specific mixed probes, the breakpoint on 1q from the derivative chromosome t(1q;7q) was identified in 1q32-q41 in esophageal KYSE140 cells. In conclusion, we established an effective labeling method for 1 Mb BAC DNA mixed clone probes, and chromosome arm and region rearrangements could be identified in several esophageal cancer cells by using these probes. Our study provides a more precise method for identification of chromosomal aberration by M-FISH, and the established method may also be applied to the karyotype analysis of hematological malignancies and prenatal diagnosis.
Jiajie Hao, Chunli Wang, Wenyue Gu, Xiaoyu Cheng, Yu Zhang, Xin Xu, Yan Cai, Mingrong Wang . Identification of chromosomal aberration in esophageal cancer cells by mixed BAC DNA probes of chromosome arms and regions[J]. Hereditas(Beijing), 2014 , 36(6) : 558 -565 . DOI: 10.3724/SP.J.1005.2014.0558
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