Decoy Wnt receptor sLRP6E1E2 inhibits cancer cell migration and invasion, and modulates expression of epithelial-to-mesenchymal transition markers and MMPs.

(a) Quantitative analysis of A549 lung cancer cell migration. Experiments were performed in triplicate, and results are expressed as mean ± SEM. *P<0.05 versus PBS- or dE1-k35/LacZ-treated controls; **P<0.001 versus PBS or dE1-k35/LacZ with Wnt3a. (b) Invasion of tumor cells was quantified as number of cells in five fields of view per filter. Experiments were performed in triplicate, and results are expressed as mean ± SEM. *P<0.05 versus PBS- or dE1-k35/LacZ-treated controls; **P<0.001 versus PBS or dE1-k35/LacZ with Wnt3a. (c) Expression of EMT markers in H322 cells after 24 hr treatment with PBS, dE1-k35/LacZ, or dE1-k35/sLRP6E1E2 in the presence and absence of Wnt3a (100 ng/ml). Cells were stained with DAPI (blue), TRITC-labeled actin (red), or anti E-cadherin (green). Original magnification, ×630. (d) Expression of EMT-related markers in H322 cell lines. Expression levels of mesenchymal markers (N-cadherin & vimentin) as well as transcriptional factor (Snail) was determined by Western blotting. (e, f) A549 cells were transduced with dE1-k35/LacZ or dE1-k35/sLRP6E1E2 with or without Wnt3a (100 ng/ml). The enzyme activity of MMP-2 and MMP-9 was measured in supernatants collected from transduced cells at 48 hr using the Sensolyte 520 MMP-2 and MMP-9 assay kit. Experiments were performed in triplicate, and results are expressed as mean ± SEM. (e) *P<0.05, (f) ^#P<0.01 versus PBS- or dE1-k35/LacZ-treated controls; (e) ^#P<0.01, (f) **P<0.001 versus PBS or dE1-k35/LacZ with Wnt3a.