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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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Vascular cell adhesion molecule-1 (VCAM-1) is involved in osteoblast-derived WISP-1-mediated PCa cell migration(A, C) PCa cells were incubated with various OBCM or WISP-1 concentrations for 24 h, and VCAM-1 expression was examined by real-time quantitative polymerase chain reaction (RT-qPCR). (B, D) PCa cells were transfected with VCAM-1 siRNA for 24 h followed by stimulation with OBCM (30 %) or WISP-1 (10 ng/mL) for 24 h; in vitro migration was measured by a Transwell assay. (E, F) Osteoblasts were transfected with control or WISP-1 shRNA for 24 h, and the medium was collected as OBCM and applied to PCa cells for 24 h. VCAM-1 expression was examined by RT-qPCR and Western blot. 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 2: Vascular cell adhesion molecule-1 (VCAM-1) is involved in osteoblast-derived WISP-1-mediated PCa cell migration(A, C) PCa cells were incubated with various OBCM or WISP-1 concentrations for 24 h, and VCAM-1 expression was examined by real-time quantitative polymerase chain reaction (RT-qPCR). (B, D) PCa cells were transfected with VCAM-1 siRNA for 24 h followed by stimulation with OBCM (30 %) or WISP-1 (10 ng/mL) for 24 h; in vitro migration was measured by a Transwell assay. (E, F) Osteoblasts were transfected with control or WISP-1 shRNA for 24 h, and the medium was collected as OBCM and applied to PCa cells for 24 h. VCAM-1 expression was examined by RT-qPCR and Western blot. Results are expressed as mean ± SEM. *, p < 0.05 compared with control; #, p < 0.05 compared with OBCM or WISP-1-treated group.

Mentions: VCAM-1 reportedly mediates tumor bone metastasis [25]. We hypothesized that VCAM-1 is involved in osteoblast-derived WISP-1-directed PCa migration. Stimulation of PCa cells with OBCM or WISP-1 increased VCAM-1 mRNA expression in a concentration-dependent manner (Fig. 2A and C). Transfection of PCa cells with VCAM-1 siRNA markedly inhibited OBCM- or WISP-1-induced migration (Fig. 2B and D). WISP-1 shRNA antagonized OBCM-mediated VCAM-1 expression (Fig. 2E–F). These data suggest that OBCM-derived WISP-1-induced PCa migration occurs via up-regulation of VCAM-1 expression. WISP-1 is known to affect cell function by binding to the cell-surface integrin receptor [30]. Incubation of PCa cells with OBCM increased mRNA expression of αv and β1 integrin (data not shown). Co-transfection of PCa cells with αvβ1 siRNA markedly reduced OBCM- or WISP-1-enhanced cell migration (Fig. 3A and D). On the other hand, αvβ1 siRNA diminished OBCM- or WISP-1-mediated VCAM-1 expression (Fig. 3B, C, E). Thus, osteoblast-derived WISP-1 increases migration and VCAM-1 expression in human PCa cells through integrin αvβ1 receptor.


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)

Vascular cell adhesion molecule-1 (VCAM-1) is involved in osteoblast-derived WISP-1-mediated PCa cell migration(A, C) PCa cells were incubated with various OBCM or WISP-1 concentrations for 24 h, and VCAM-1 expression was examined by real-time quantitative polymerase chain reaction (RT-qPCR). (B, D) PCa cells were transfected with VCAM-1 siRNA for 24 h followed by stimulation with OBCM (30 %) or WISP-1 (10 ng/mL) for 24 h; in vitro migration was measured by a Transwell assay. (E, F) Osteoblasts were transfected with control or WISP-1 shRNA for 24 h, and the medium was collected as OBCM and applied to PCa cells for 24 h. VCAM-1 expression was examined by RT-qPCR and Western blot. 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 2: Vascular cell adhesion molecule-1 (VCAM-1) is involved in osteoblast-derived WISP-1-mediated PCa cell migration(A, C) PCa cells were incubated with various OBCM or WISP-1 concentrations for 24 h, and VCAM-1 expression was examined by real-time quantitative polymerase chain reaction (RT-qPCR). (B, D) PCa cells were transfected with VCAM-1 siRNA for 24 h followed by stimulation with OBCM (30 %) or WISP-1 (10 ng/mL) for 24 h; in vitro migration was measured by a Transwell assay. (E, F) Osteoblasts were transfected with control or WISP-1 shRNA for 24 h, and the medium was collected as OBCM and applied to PCa cells for 24 h. VCAM-1 expression was examined by RT-qPCR and Western blot. Results are expressed as mean ± SEM. *, p < 0.05 compared with control; #, p < 0.05 compared with OBCM or WISP-1-treated group.
Mentions: VCAM-1 reportedly mediates tumor bone metastasis [25]. We hypothesized that VCAM-1 is involved in osteoblast-derived WISP-1-directed PCa migration. Stimulation of PCa cells with OBCM or WISP-1 increased VCAM-1 mRNA expression in a concentration-dependent manner (Fig. 2A and C). Transfection of PCa cells with VCAM-1 siRNA markedly inhibited OBCM- or WISP-1-induced migration (Fig. 2B and D). WISP-1 shRNA antagonized OBCM-mediated VCAM-1 expression (Fig. 2E–F). These data suggest that OBCM-derived WISP-1-induced PCa migration occurs via up-regulation of VCAM-1 expression. WISP-1 is known to affect cell function by binding to the cell-surface integrin receptor [30]. Incubation of PCa cells with OBCM increased mRNA expression of αv and β1 integrin (data not shown). Co-transfection of PCa cells with αvβ1 siRNA markedly reduced OBCM- or WISP-1-enhanced cell migration (Fig. 3A and D). On the other hand, αvβ1 siRNA diminished OBCM- or WISP-1-mediated VCAM-1 expression (Fig. 3B, C, E). Thus, osteoblast-derived WISP-1 increases migration and VCAM-1 expression in human PCa cells through integrin αvβ1 receptor.

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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Related in: MedlinePlus