The frequency of osteolytic bone metastasis is determined by conditions of the soil, not the number of seeds; evidence from in vivo models of breast and prostate cancer.
BACKGROUND
While both preclinical and clinical studies suggest that the frequency of growing skeletal metastases is elevated in individuals with higher bone turnover, it is unclear whether this is a result of increased numbers of tumour cells arriving in active sites or of higher numbers of tumour cells being induced to divide by the bone micro-environment. Here we have investigated how the differences in bone turnover affect seeding of tumour cells and/or development of overt osteolytic bone metastasis using in vivo models of hormone-independent breast and prostate cancer.


METHODS
Cohorts of 6 (young) and 16 (mature)-week old BALB/c nude mice were culled 1, 7 and 21 days after received intracardiac injection of luciferase expressing human prostate (PC3) or breast cancer (MDA-MB-231) cell lines labelled with a fluorescent cell membrane dye (Vybrant DiD). The presence of growing bone metastases was determined by bioluminescence using an in vivo imaging system (IVIS) and followed by anatomical confirmation of tumour metastatic sites post mortem, while the presence of individual fluorescently labelled tumour cells was evaluated using two-photon microscopy ex vivo. The bone remodelling activities were compared between young and mature naïve mice (both male and female) using micro-CT analysis, ELISA and bone histomorphometry.


RESULTS
Both prostate and breast cancer cells generated higher numbers of overt skeletal lesions in young mice (~80%) than in mature mice (~20%). Although mature mice presented with fewer overt bone metastases, the number of tumour cells arriving/colonizing in the tibias was comparable between young and mature animals. Young naïve mice had lower bone volume but higher bone formation and resorption activities compared to mature animals.


CONCLUSIONS
Our studies suggest that higher frequencies of growing osteolytic skeletal metastases in these models are linked to increased bone turnover and not to the initial number of tumour cells entering the bone microenvironment.
The Mellanby Centre for Bone Research, Department of Human Metabolism, Medical School, University of Sheffield, Beech Hill Road, Sheffield, S10 2RX, UK. n.wang@sheffield.ac.uk.
Ning
N
Wang
Break Through Breast Cancer Research Unit, Paterson Institute for Cancer Research Manchester, Manchester, UK. kimberley.reeves@manchester.ac.uk.
Kimberley J
KJ
Reeves
The Mellanby Centre for Bone Research, Department of Human Metabolism, Medical School, University of Sheffield, Beech Hill Road, Sheffield, S10 2RX, UK. hannah.brown@sheffield.ac.uk.
Hannah K
HK
Brown
The Mellanby Centre for Bone Research, Department of Human Metabolism, Medical School, University of Sheffield, Beech Hill Road, Sheffield, S10 2RX, UK. a.m.fowles@sheffield.ac.uk.
Anne C M
AC
Fowles
The Mellanby Centre for Bone Research, Department of Human Metabolism, Medical School, University of Sheffield, Beech Hill Road, Sheffield, S10 2RX, UK. freyjadocherty@hotmail.co.uk.
Freyja E
FE
Docherty
Department of Oncology, Medical School, University of Sheffield, Sheffield, UK. P.D.Ottewell@sheffield.ac.uk.
Penelope D
PD
Ottewell
Bone Biology Division, Garvan Institute of Medical Research, Sydney, Australia. p.croucher@garvan.org.au.
Peter I
PI
Croucher
Department of Oncology, Medical School, University of Sheffield, Sheffield, UK. i.holen@sheffield.ac.uk.
Ingunn
I
Holen
The Mellanby Centre for Bone Research, Department of Human Metabolism, Medical School, University of Sheffield, Beech Hill Road, Sheffield, S10 2RX, UK. c.l.eaton@sheffield.ac.uk.
Colby L
CL
Eaton
D000818:Animals; D001859:Bone Neoplasms / Q000473:pathology* / Q000556:secondary; D001943:Breast Neoplasms / Q000473:pathology*; D002452:Cell Count; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D005260:Female; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D010014:Osteolysis / Q000473:pathology*; D011471:Prostatic Neoplasms / Q000473:pathology*; D014183:Transplantation, Heterologous