Skip to main content
. 2016 Oct 20;7(47):77071–77086. doi: 10.18632/oncotarget.12787

Figure 4. ERRα overexpression in PCa-ERRα cells stimulates bone-forming cells.

Figure 4

(A) Osteoblasts (Ob) from primary mouse calvaria cell cultures were treated with conditioned medium of PC3-ERRα (ERRα) or PC3-CT (CT) cells. Bone nodule number was higher in co-cultures with PC3-overexpressing ERRα (Ob-ERRα) (Mann-Whitney,P = 0.0118). (B) When treated with conditioned medium of PC3c-ERRα (ERRα), bone nodule number was increased but not significantly compared to PC3c-CT (CT). (C) Confirming mouse calvaria results, co-cultures of PC3-ERRα (ERRα) or PC3-CT (CT) cells with the MC3T3-E1 osteoblastic cell line for 2 weeks stimulated expression of osteoblastic markers alp (Alcalin Phosphatase), bsp (bone sialoprotein) and ocn (osteocalcin) (ANOVA, P < 0.0001; Student's t-tests, P = 0.0031; P = 0.0098; P = 0.0017) (D) Expression of rankl mRNA was also higher (ANOVA, P < 0.0001, Student's t-tests, P = 0.0279) while opg expression was not altered. (E) Co-cultures of ACE-1-ERRα (ERRα) or ACE-1-CT (CT) cells with the MC3T3-E1 osteoblastic cell line for 2 weeks also stimulated bsp and ocn mRNA expression (ANOVA bsp, P = 0.004; Student's t-tests, P = 0.0003 (ACE-1-CT versus ACE-1-ERRα); ANOVA ocn, P = 0.0069; Student's t-tests, P = 0.0067 (ACE-1-CT versus ACE-1-ERRα) while no regulation was observed with ACE-1-AF2 cells (AF2). (F) Expression of rankl and opg mRNA was not regulated. Ob results are representative of two independent experiments, each performed in triplicate samples.