Stimulation of TLR4 by LMW-HA Induces Metastasis in Human Papillary Thyroid Carcinoma through CXCR7
Figure 2
LMW-HA promotes W3 cell proliferation and migration via TLR4. (a) W3 cells were seeded into 96-well plates (2,000 cells/well) and treated with or without LMW-HA. Cell proliferation was analyzed with WST-1 Kit. Data are mean ± SEM for three independent experiments. (b) W3 cells were treated with or without LMW-HA for 24 hours and stained with annexin V and PI. Data are mean ± SEM for three independent experiments. (c) W3 cells were seeded into the upper chambers of transwell inserts treated with or without LMW-HA and in the presence or absence of CXCL12 in the lower chambers. Migrated cells were determined. Data are mean ± SEM for three independent experiments. (d) W3 cells were transfected with scrambled shRNA (Scram) or TLR4 shRNA-expressing constructs (TLR4-KD) and subjected to immunoblot analysis. (e) Scram-W3 cells and TLR4-KD W3 cells were seeded into 96 well plates and treated with or without LMW-HA (100 μg/mL) for 24 h; cell proliferation was analyzed. Data are mean ± SEM for three independent experiments. (f) Scram-W3 cell and TLR4-KD W3 cell migration to CXCL12 treated with or without LMW-HA was determined. Data are mean ± SEM for three independent experiments.
