Application of the polymerase chain reaction (PCR) to quantify micro-metastasis in an experimental animal.
In vitro cultured r/mHM-SFME-1 cells were injected into the hind foot pads of Balb/c mice. Metastasis was detected in the lungs of tumor-bearing mice by means of both PCR and histological methods. Primers for the PCR were set to amplify a 128 bp exon-1 sequence of the human c-Ha-ras1 gene which had been introduced into the cells. Resulted PCR bands were densitometorically quantified using a bioimage analyzer, and more than 1 x 10(4) tumor cells were detectable in the mouse lung. The number of tumor cells per lung estimated from the amount of PCR products was 1 x 10(5), 15 x 10(5), 1 x 10(5) and 40 x 10(5) on days 7, 14, 21 and 28 respectively after the tumor injection. No metastases were histologically observed on days 7 and 14. Then, the possibility of using this model system for evaluation of a treatment against micro-metastases is discussed.
Matano
S
S
Department of 3rd Intern. Med., School Med., Kanazawa Univ., Ishikawa, Japan.
Ryoyama
K
K
Nakamura
S
S
Okada
G
G
Nomura
T
T
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002461:Cell Line, Transformed; D011905:Genes, ras; D008175:Lung Neoplasms / Q000235:genetics / Q000473:pathology / Q000556:secondary*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009362:Neoplasm Metastasis / Q000235:genetics* / Q000473:pathology; D016133:Polymerase Chain Reaction