Figure 2.

COL6A1 promotes migration and invasion in vivo and in vitro. A. Overexpression of COL6A1 increased OS cell migration and invasive abilities detected by transwell assay in OS cell lines, U2OS and Saos-2. B. Downregulation of COL6A1 by siRNA transfection resulted in a decrease in the migratory and invasive abilities of OS cells as determined by transwell analysis. C. The migration ability of OS cells was detected by wound-healing assay after COL6A1 or siCOL6A1 transfection. D. Anoikis assay: Cell viability determined by MTT in COL6A1 overexpressed, knockdown or control cells seeded at 10⁵ cells/well in a 96 poly-Hema culture plate. E. COL6A1-overexpressing, COL6A1 knockdown or control cells were subjected to cell-matrix adhesion assay to collagen I (Col I), collagen IV (Col IV), laminin (LN), and fibronectin (FN). F. Western blotting analysis of phosphorylation of FAK and Src and total FAK and Src in COL6A1-overexpressing and control cells upon treatment with vehicle or Src inhibitor (10 µM) for 24 h. G. COL6A1-overexpressing and control cells were subjected to cell-matrix adhesion assay to Col I, Col IV, and FN upon treatment with vehicle or Src inhibitor (10 µM) for 24 h. H. Cell migration potential was determined in COL6A1-overexpressing and control cells upon treatment with vehicle or Src inhibitor for 24 h according to transwell assays. I. Overexpression of COL6A1 increased the rate of lung metastasis after tail-vein injection of COL6A1 or control cells (n = 6 each group). Representative photographs of hematoxylin and eosin (H&E) staining in lung metastases tissues from mice orthotopically inoculated with COL6A1 and control cells (Scale bar: 200 µm, 50 µm). Data represent the mean ± SD of 3 separate determinations. *p < 0.05, **p < 0.01, ***p < 0.001 by Student's t test.