Bone
metastasis from breast and prostate
carcinomas is facilitated by activation of bone-resorbing osteoclasts. Using proteomics approaches, we have identified
peroxiredoxin-4 (PRDX4) as a
cancer-secreted mediator of osteoclastogenesis. We now report characterization of
L-plastin in the
conditioned media (CM) of MDA-MB-231 human
breast cancer cells using immunoblotting and mass spectrometry. The osteoclastogenic potential of MDA-MB-231 CM with
siRNA-silenced
L-plastin was significantly reduced.
L-plastin was detected in
cancer-derived exosomes, and inhibition of exosomal release significantly decreased the osteoclastogenic capacity of MDA-MB-231 CM. When added to osteoclast precursors primed with RANKL for 2 days, recombinant
L-plastin induced
calcium/NFATc1-mediated osteoclastogenesis to the levels similar to continuous treatment with RANKL. Using
shRNA, we generated MDA-MB-231 cells lacking
L-plastin, PRDX4, or both and injected these cell populations intratibially in CD-1 immunodeficient mice. Micro-CT and histomorphometric analysis demonstrated a complete loss of
osteolysis when MDA-MB-231 cells lacking both
L-plastin and PRDX4 were injected. A meta-analysis established an increase in
L-plastin and PRDX4
mRNA expression in numerous human
cancers, including breast and prostate
carcinomas. This study demonstrates that secreted
L-plastin and PRDX4 mediate osteoclast activation by human
breast cancer cells.