TY - JOUR
T1 - Identification of novel breakpoints for locus- and region-specific translocations in 293 cells by molecular cytogenetics before and after irradiation
AU - Binz, Regina L.
AU - Tian, Erming
AU - Sadhukhan, Ratan
AU - Zhou, Daohong
AU - Hauer-Jensen, Martin
AU - Pathak, Rupak
N1 - Funding Information:
This manuscript was edited by the Science Communication Group at the University of Arkansas for Medical Sciences (UAMS). The authors acknowledge Ms. Sandra Avra (Molecular Genetics Lab, UAMS Pathology) for proofreading the nomenclature. This study was supported by Arkansas Space Grant Consortium grant NNX15AR71H (R.P.); an Institutional Development Award (IDeA) from the NIGMS of the National Institutes of Health under grant number P20 GM109005 (R.P. and M.H.J.), and National Cancer Institute grant 7R01CA211963-02 (R.P. and D.Z.). This manuscript was edited by the Science Communication Group at the University of Arkansas for Medical Sciences (UAMS).
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - The human kidney embryonic 293 cell line (293 cells) is extensively used in biomedical and pharmaceutical research. These cells exhibit a number of numerical and structural chromosomal anomalies. However, the breakpoints responsible for these structural chromosomal rearrangements have not been comprehensively characterized. In addition, it is not known whether chromosomes with structural rearrangement are more sensitive to external toxic agents, such as ionizing radiation. We used G-banding, spectral karyotyping (SKY), and locus- and region-specific fluorescence in situ hybridization (FISH) probes designed in our lab or obtained from commercial vendor to address this gap. Our G-banding analysis revealed that the chromosome number varies from 66 to 71, with multiple rearrangements and partial additions and deletions. SKY analysis confirmed 3 consistent rearrangements, two simple and one complex in nature. Multicolor FISH analysis identified an array of breakpoints responsible for locus- and region-specific translocations. Finally, SKY analysis revealed that radio-sensitivity of structurally rearranged chromosomes is dependent on radiation dose. These findings will advance our knowledge in 293 cell biology and will enrich the understanding of radiation biology studies.
AB - The human kidney embryonic 293 cell line (293 cells) is extensively used in biomedical and pharmaceutical research. These cells exhibit a number of numerical and structural chromosomal anomalies. However, the breakpoints responsible for these structural chromosomal rearrangements have not been comprehensively characterized. In addition, it is not known whether chromosomes with structural rearrangement are more sensitive to external toxic agents, such as ionizing radiation. We used G-banding, spectral karyotyping (SKY), and locus- and region-specific fluorescence in situ hybridization (FISH) probes designed in our lab or obtained from commercial vendor to address this gap. Our G-banding analysis revealed that the chromosome number varies from 66 to 71, with multiple rearrangements and partial additions and deletions. SKY analysis confirmed 3 consistent rearrangements, two simple and one complex in nature. Multicolor FISH analysis identified an array of breakpoints responsible for locus- and region-specific translocations. Finally, SKY analysis revealed that radio-sensitivity of structurally rearranged chromosomes is dependent on radiation dose. These findings will advance our knowledge in 293 cell biology and will enrich the understanding of radiation biology studies.
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U2 - 10.1038/s41598-019-47002-0
DO - 10.1038/s41598-019-47002-0
M3 - Article
C2 - 31332273
AN - SCOPUS:85070250547
SN - 2045-2322
VL - 9
JO - Scientific Reports
JF - Scientific Reports
IS - 1
M1 - 10554
ER -