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Osteoblast-derived WNT-induced secreted protein 1 increases VCAM-1 expression and enhances prostate cancer metastasis by down-regulating miR-126.

Tai HC, Chang AC, Yu HJ, Huang CY, Tsai YC, Lai YW, Sun HL, Tang CH, Wang SW - Oncotarget (2014)

Bottom Line: Osteoblast transfection with WISP-1 shRNA reduced OBCM-mediated PCa migration and VCAM-1 expression.Osteoblast-derived WISP-1 inhibited miR-126 expression.This study suggests that osteoblast-derived WISP-1 promotes migration and VCAM-1 expression in human PCa cells by down-regulating miR-126 expression via αvβ1 integrin, FAK, and p38 signaling pathways.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, National Taiwan University Hospital, Taipei, Taiwan.

ABSTRACT
Bone metastases of prostate cancer (PCa) may cause intractable pain. Wnt-1-induced secreted protein 1 (WISP-1) belongs to the CCN family (CTGF/CYR61/NOV) that plays a key role in bone formation. We found that osteoblast-conditioned medium (OBCM) stimulates migration and vascular adhesion molecule-1 (VCAM)-1 expression in human PCa (PC3 and DU145) cells. Osteoblast transfection with WISP-1 shRNA reduced OBCM-mediated PCa migration and VCAM-1 expression. Stimulation of PCa with OBCM or WISP-1 elevated focal adhesion kinase (FAK) and p38 phosphorylation. Either FAK and p38 inhibitors or siRNA abolished osteoblast-derived WISP-1-induced migration and VCAM-1 expression. Osteoblast-derived WISP-1 inhibited miR-126 expression. Moreover, miR-216 mimic reversed the WISP-1-enhanced migration and VCAM-1 expression. This study suggests that osteoblast-derived WISP-1 promotes migration and VCAM-1 expression in human PCa cells by down-regulating miR-126 expression via αvβ1 integrin, FAK, and p38 signaling pathways. Thus, WISP-1 may be a new molecular therapeutic target in PCa bone metastasis.

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FAK and p38 pathways are involved in osteoblast-derived WISP-1-increased migration and VCAM-1 expression(A) Osteoblasts were transfected with control or WISP-1 shRNA for 24 h, and the medium was collected as OBCM and applied to PCa cells. FAK, p38, ERK, and JNK phosphorylation was examined by Western blot. (B) PCa cells were incubated with WISP-1 (10 ng/mL) for the indicated time intervals; FAK and p38 phosphorylation was examined by Western blot. (C–H) PCa cells were pretreated with FAK inhibitor (10 μM) and SB203580 (10 μM) or transfected with FAK and p38 siRNA for 24 h followed by stimulation with OBCM (30 %) or WISP-1 (10 ng/mL) for 24 h; in vitro migration and VCAM-1 expression were measured by Transwell, RT-qPCR, and Western blot analyses. Results are expressed as mean ± SEM. *, p < 0.05 compared with control; #, p < 0.05 compared with OBCM or WISP-1-treated group.
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Figure 4: FAK and p38 pathways are involved in osteoblast-derived WISP-1-increased migration and VCAM-1 expression(A) Osteoblasts were transfected with control or WISP-1 shRNA for 24 h, and the medium was collected as OBCM and applied to PCa cells. FAK, p38, ERK, and JNK phosphorylation was examined by Western blot. (B) PCa cells were incubated with WISP-1 (10 ng/mL) for the indicated time intervals; FAK and p38 phosphorylation was examined by Western blot. (C–H) PCa cells were pretreated with FAK inhibitor (10 μM) and SB203580 (10 μM) or transfected with FAK and p38 siRNA for 24 h followed by stimulation with OBCM (30 %) or WISP-1 (10 ng/mL) for 24 h; in vitro migration and VCAM-1 expression were measured by Transwell, RT-qPCR, and Western blot analyses. Results are expressed as mean ± SEM. *, p < 0.05 compared with control; #, p < 0.05 compared with OBCM or WISP-1-treated group.

Mentions: FAK, a widely expressed non-receptor protein tyrosine kinase, is an early downstream factor of integrin-mediated signaling that regulates cellular function [31]. To verify whether FAK activation is involved in osteoblast-derived WISP-1-induced cell migration, we directly measured phosphorylation of FAK in response to OBCM. Stimulation of PCa cells with OBCM increased FAK phosphorylation (Fig. 4A). In contrast, knockdown of WISP-1 in osteoblasts diminished OBCM-mediated FAK phosphorylation (Fig. 4A). FAK inhibitor or siRNA reduced OBCM- or WISP-1-increased cell migration and VCAM-1 expression in human PCa (Fig. 4C–H), indicating that WISP-1 enhanced FAK phosphorylation (Fig. 4B).


Osteoblast-derived WNT-induced secreted protein 1 increases VCAM-1 expression and enhances prostate cancer metastasis by down-regulating miR-126.

Tai HC, Chang AC, Yu HJ, Huang CY, Tsai YC, Lai YW, Sun HL, Tang CH, Wang SW - Oncotarget (2014)

FAK and p38 pathways are involved in osteoblast-derived WISP-1-increased migration and VCAM-1 expression(A) Osteoblasts were transfected with control or WISP-1 shRNA for 24 h, and the medium was collected as OBCM and applied to PCa cells. FAK, p38, ERK, and JNK phosphorylation was examined by Western blot. (B) PCa cells were incubated with WISP-1 (10 ng/mL) for the indicated time intervals; FAK and p38 phosphorylation was examined by Western blot. (C–H) PCa cells were pretreated with FAK inhibitor (10 μM) and SB203580 (10 μM) or transfected with FAK and p38 siRNA for 24 h followed by stimulation with OBCM (30 %) or WISP-1 (10 ng/mL) for 24 h; in vitro migration and VCAM-1 expression were measured by Transwell, RT-qPCR, and Western blot analyses. Results are expressed as mean ± SEM. *, p < 0.05 compared with control; #, p < 0.05 compared with OBCM or WISP-1-treated group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4202146&req=5

Figure 4: FAK and p38 pathways are involved in osteoblast-derived WISP-1-increased migration and VCAM-1 expression(A) Osteoblasts were transfected with control or WISP-1 shRNA for 24 h, and the medium was collected as OBCM and applied to PCa cells. FAK, p38, ERK, and JNK phosphorylation was examined by Western blot. (B) PCa cells were incubated with WISP-1 (10 ng/mL) for the indicated time intervals; FAK and p38 phosphorylation was examined by Western blot. (C–H) PCa cells were pretreated with FAK inhibitor (10 μM) and SB203580 (10 μM) or transfected with FAK and p38 siRNA for 24 h followed by stimulation with OBCM (30 %) or WISP-1 (10 ng/mL) for 24 h; in vitro migration and VCAM-1 expression were measured by Transwell, RT-qPCR, and Western blot analyses. Results are expressed as mean ± SEM. *, p < 0.05 compared with control; #, p < 0.05 compared with OBCM or WISP-1-treated group.
Mentions: FAK, a widely expressed non-receptor protein tyrosine kinase, is an early downstream factor of integrin-mediated signaling that regulates cellular function [31]. To verify whether FAK activation is involved in osteoblast-derived WISP-1-induced cell migration, we directly measured phosphorylation of FAK in response to OBCM. Stimulation of PCa cells with OBCM increased FAK phosphorylation (Fig. 4A). In contrast, knockdown of WISP-1 in osteoblasts diminished OBCM-mediated FAK phosphorylation (Fig. 4A). FAK inhibitor or siRNA reduced OBCM- or WISP-1-increased cell migration and VCAM-1 expression in human PCa (Fig. 4C–H), indicating that WISP-1 enhanced FAK phosphorylation (Fig. 4B).

Bottom Line: Osteoblast transfection with WISP-1 shRNA reduced OBCM-mediated PCa migration and VCAM-1 expression.Osteoblast-derived WISP-1 inhibited miR-126 expression.This study suggests that osteoblast-derived WISP-1 promotes migration and VCAM-1 expression in human PCa cells by down-regulating miR-126 expression via αvβ1 integrin, FAK, and p38 signaling pathways.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, National Taiwan University Hospital, Taipei, Taiwan.

ABSTRACT
Bone metastases of prostate cancer (PCa) may cause intractable pain. Wnt-1-induced secreted protein 1 (WISP-1) belongs to the CCN family (CTGF/CYR61/NOV) that plays a key role in bone formation. We found that osteoblast-conditioned medium (OBCM) stimulates migration and vascular adhesion molecule-1 (VCAM)-1 expression in human PCa (PC3 and DU145) cells. Osteoblast transfection with WISP-1 shRNA reduced OBCM-mediated PCa migration and VCAM-1 expression. Stimulation of PCa with OBCM or WISP-1 elevated focal adhesion kinase (FAK) and p38 phosphorylation. Either FAK and p38 inhibitors or siRNA abolished osteoblast-derived WISP-1-induced migration and VCAM-1 expression. Osteoblast-derived WISP-1 inhibited miR-126 expression. Moreover, miR-216 mimic reversed the WISP-1-enhanced migration and VCAM-1 expression. This study suggests that osteoblast-derived WISP-1 promotes migration and VCAM-1 expression in human PCa cells by down-regulating miR-126 expression via αvβ1 integrin, FAK, and p38 signaling pathways. Thus, WISP-1 may be a new molecular therapeutic target in PCa bone metastasis.

Show MeSH
Related in: MedlinePlus