Specific detection of metastasized human tumor cells in embryonic chicks by the polymerase chain reaction.
We have established a highly sensitive method for specific detection of metastasized human tumor cells in embryonic chicks using the polymerase chain reaction (PCR). The cells (HT-1080 or KMST-6) were inoculated into the chorioallantoic membrane vein of the chick embryo and DNA of each embryonic organ was extracted. Then, a human beta-globin-related sequence (576 bp) in the DNA from the embryonic liver and lung was specifically amplified and detected by gel electrophoresis and by a specific oligonucleotide probe. The amplified fragments from the liver DNA samples increased gradually from 2 h to 7 days after HT-1080 inoculation. On the other hand, with inoculation of non-tumorigenic human embryonal fibroblast KMST-6 cells, the DNA from the embryonic liver 7 days after inoculation did not show the PCR-amplified product. This detection technique can contribute significantly to the precise detection of microscopic metastasis.
Endo
Y
Y
Department of Experimental Therapeutics, Kanazawa University.
Sasaki
T
T
Harada
F
F
Noguchi
M
M
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002452:Cell Count; D002642:Chick Embryo; D004273:DNA, Neoplasm / Q000032:analysis*; D005354:Fibrosarcoma / Q000235:genetics / Q000473:pathology / Q000556:secondary; D005784:Gene Amplification / Q000235:genetics; D006801:Humans; D008099:Liver / Q000032:analysis / Q000473:pathology; D008969:Molecular Sequence Data; D009362:Neoplasm Metastasis / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction / Q000379:methods*; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm