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Results: 4

1.
Figure 3

Figure 3. From: Wnt Inhibitory Factor 1 (WIF-1) decreases tumorigenesis and metastasis in osteosarcoma.

WIF-1 inhibits in vivo tumor growth in nude mice. 1 × 106 143B cells per mice were inoculated subcutaneously into the left flank of NCR nu-nu (nude) mice. A, Tumor growth curve. Points, mean tumor volume (each group contains 10 mice); Bars, standard error (SE). B, Tumor wet weight. Tumors were harvested at 18 days after inoculation. Columns, mean wet tumor weight from 10 mice in each group; Bars, SE. A representative photograph of harvested tumors is shown (insert)

Elyssa M. Rubin, et al. Mol Cancer Ther. ;9(3):731-741.
2.
Figure 1

Figure 1. From: Wnt Inhibitory Factor 1 (WIF-1) decreases tumorigenesis and metastasis in osteosarcoma.

WIF-1 expression is down-regulated in OS cell lines, which is associated with promoter methylation. A, mRNA levels of WIF-1 in 7 OS cell lines (SaOS-2, SaOS-LM7, MNNG-HOS, 143B, 143.98.2, U2-OS, OS-160) and normal human osteoblasts (NHOst) were determined by real-time RT-PCR. After all these osteosarcoma cell lines and normal osteoblast were treated with 5-Aza-2′-deoxycytidine (5-Aza-dC; 5 μmol/L) for 72 h, real time RT-PCR were performed to detect the WIF-1 mRNA level. Experiments were replicated three times. B, WIF-1 promoter hypermethylation is evident in 5 of 7 OS cell lines. Methylation status of WIF-1 promoter was determined by disulfite treatment of cell lines followed by methylation-specific PCR. C, Expression of WIF-1 proteins were detected by Western blottingting with an anti-hWIF-1 antibody. D, Representative photograph (200X magnification) of negative versus positive immunohistochemical staining of human OS tissue microarray using an anti-hWIF-1 antibody.

Elyssa M. Rubin, et al. Mol Cancer Ther. ;9(3):731-741.
3.
Figure 4

Figure 4. From: Wnt Inhibitory Factor 1 (WIF-1) decreases tumorigenesis and metastasis in osteosarcoma.

WIF-1 reduces lung metastasis in an orthotopic OS mouse model and down-regulated the expression of MMP-9 and 14. A, lungs from mice injected with WIF-1 and vector control transfected 143B cells were harvested and fixed in Bouin’s solution on day 21. Surface lung metastatic nodules were pointed out by arrows. B, Surface lung nodules were counted under a dissecting microscope. Columns, mean number of lung surface nodules from 10 mice in each group; Bars, SE. A representative photograph of lungs from each transfection group is shown (insert). C, representative photographs of lung sections from WIF-1 and vector control transfection groups by H&E staining (×40 magnification). Lung metastatic nodules were pointed out by arrows. D, Protein expression of WIF-1, MMP-9 and MMP-14 in WIF-1 and vector control transfected 143B cell lines was determined by Western blotting assay.

Elyssa M. Rubin, et al. Mol Cancer Ther. ;9(3):731-741.
4.
Figure 2

Figure 2. From: Wnt Inhibitory Factor 1 (WIF-1) decreases tumorigenesis and metastasis in osteosarcoma.

Ectopic WIF-1 expression inhibits colony formation in soft agar and cell migration of 143B cells. A, WIF-1 transfection reduced TCF4 transcriptional activity in 143B cells compared to vector control transfected cells, measured by TOPFLASH reporter assay and adjusted by transfection efficiency. The expression of V5-tagged WIF-1 in transfected 143B cells was confirmed by Western blotting using an anti-V5 antibody (insert). B, Anchorage-independent colony formation assay showed decreased number of colonies formed by WIF-1 transfection compared to vector control transfection of 143B cells; A representative photograph of soft agar colonies at 18 days after cell seeding (insert). C, Representative photomicrographs of scratch wounds at 0 hours and 12 hours after wounds were made. D, Quantitative measurement of wound gaps by Photoshop® software showed a reduced cellular motility in the WIF-1 transfected 143B cell line compared to vector control cells. Columns represent mean relative change (%) of wound width in WIF-1 vs vector control transfected 143B cells. Bars, standard deviation (SD). Experiments were replicated three times.

Elyssa M. Rubin, et al. Mol Cancer Ther. ;9(3):731-741.

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