The Effects of a Novel Hormonal Breast Cancer Therapy, Endoxifen, on the Mouse Skeleton
Figure 7
Cellular responses to vehicle and endoxifen treatment.
A. Real-time PCR analysis of alkaline phosphatase (AP), osterix (OX), Runx2 (RX2), estrogen receptor α (ERα) and estrogen receptor β (ERβ) in adherent marrow stromal cells derived from endoxifen treated mice relative to vehicle treated control animals. B. Real-time PCR analysis of AP, OX, and RX2 following 24 hour treatment of human fetal osteoblast cells expressing ERα (FOB/ER9) with 100 nM or 1000 nM levels of endoxifen. C. Real-time PCR analysis of AP, OX, RX2, ERα, ERβ, matrix extracellular phosphoglycoprotein (MEPE), phosphate-regulated neutral endopeptidase (PHEX) and dentin matrix acidic phosphoprotein1 (DMP1) in cortical shells isolated from endoxifen treated mice relative to vehicle treated control animals. D. Quantification of TRAP positive osteoclasts (OC) after MCSF and RANKL treatment of non-adherent bone marrow cells isolated from vehicle (Veh) or endoxifen (End) treated mice. E. A representative image of differentiated osteoclasts from vehicle and endoxifen treated mice. F. RT-PCR analysis of the osteoclast marker genes NFATc1, RANK, c-Fms and CathK, as well as the inhibitory OCIL gene, in mature osteoclasts derived from endoxifen treated animals relative to vehicle treated controls. The mean ± SE are depicted. * denotes significance at P<0.05.