title: Human breast cancer cell metastasis to long bone and soft organs of nude mice: a quantitative assay.
abstract: Bone is a common metastatic site in human breast cancer (HBC). Since bone metastasis occurs very rarely from current spontaneous or experimental metastasis models of HBC cells in nude mice, an arterial seeding model involving the direct injection of the cells into the left ventricle has been developed to better understand the mechanisms involved in this process. We present here a sensitive polymerase chain reaction (PCR) method to detect and quantitate bone and soft organ metastasis in nude mice which have been intracardially inoculated with Lac Z transduced HBC cells. Amplification of genomically incorporated Lac Z sequences in MDA-MB-231-BAG HBC cells enables us to specifically detect these cells in mouse organs and bones. We have also created a competitive template to use as an internal standard in the PCR reactions, allowing us to better quantitate levels of HBC metastasis. The results of this PCR detection method correlate well with cell culture detection from alternate long bones from the same mice, and are more sensitive than gross Lac Z staining with X-gal or routine histology. Comparable qualitative results were obtained with PCR and culture in a titration experiment in which mice were inoculated with increasing numbers of cells, but PCR is more quantifiable, less time consuming, and less expensive. This assay can be employed to study the molecular and cellular aspects of bone metastasis, and could easily be used in conjunction with RT-PCR-based analyses of gene products which may be involved with HBC metastasis.
authors.affiliation: Lombardi Cancer Center and Department of Cell Biology, Georgetown University Medical Center, Washington, DC 20007, USA.
authors.forename: V
authors.initials: V
authors.lastname: Sung
authors.forename: D A
authors.initials: DA
authors.lastname: Cattell
authors.forename: J M
authors.initials: JM
authors.lastname: Bueno
authors.forename: A
authors.initials: A
authors.lastname: Murray
authors.forename: J A
authors.initials: JA
authors.lastname: Zwiebel
authors.forename: A D
authors.initials: AD
authors.lastname: Aaron
authors.forename: E W
authors.initials: EW
authors.lastname: Thompson
mesh_terms: D000818:Animals; D001859:Bone Neoplasms / Q000556:secondary*; D001943:Breast Neoplasms / Q000235:genetics / Q000473:pathology*; D002452:Cell Count; D005260:Female; D017930:Genes, Reporter; D006801:Humans; D007763:Lac Operon; D051379:Mice; D008819:Mice, Nude; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012983:Soft Tissue Neoplasms / Q000556:secondary*; D014407:Tumor Cells, Cultured