A genetically engineered V79 cell line SD1 expressing rat CYP2B1 exhibits chromosomal instability at the integration site of the transfected DNA.
The genetically engineered cell line SD1 was constructed by co-transfection of V79 Chinese hamster cells with two plasmids: one containing a full-length cDNA encoding rat CYP2B1 and the second incorporating a selective marker gene. This cell line has been used in gene mutation tests and in cytokinesis-block micronucleus assays to identify procarcinogens which are metabolized by CYP2B1 to reactive metabolites. An elevated frequency of spontaneous micronuclei was recorded in SD1 cells compared to parental V79 cultures. Karyotypic analysis revealed a chromosomal instability which was manifested by amplification of the p-arms of a chromosome designated 'n' (derived from chromosome 8). This chromosome was variable in length and sometimes exhibited a telomeric fusion which led to the formation of a dicentric chromosome. Fluorescence in situ hybridization with digoxigenin-labelled plasmid DNA showed the presence of pSV450 plasmid DNA coamplified with genomic DNA sequences located in the terminal region of chromosome 'n'.
Ellard
S
S
School of Biological Sciences, University of Wales, Singleton Park, Swansea, UK.
James
S A
SA
Parry
E M
EM
Parry
J M
JM
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D002843:Chromatin / Q000235:genetics; D002869:Chromosome Aberrations; D002871:Chromosome Banding; D006224:Cricetinae; D003412:Cricetulus; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D004076:Digoxigenin / Q000737:chemistry; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology*; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D008168:Lung / Q000648:ultrastructure; D015162:Micronucleus Tests; D008938:Mitosis; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D011123:Polyploidy; D051381:Rats; D008941:Spindle Apparatus / Q000648:ultrastructure; D014162:Transfection / Q000235:genetics*
D002843:Chromatin; D003577:Cytochrome P-450 Enzyme System; D004076:Digoxigenin