Journal:

Article Title: Furin inhibition results in absent or decreased invasiveness and tumorigenicity of human cancer cells

doi: 10.1073/pnas.191199198

Figure Lengend Snippet: (A) Western blot analysis of furin expression in HNSCC. Moderate to high levels of furin were seen in four cell lines. Detroit 562 and A 253 are aggressive cell lines that showed high levels of invasiveness in vivo, whereas SCC 15 and SCC 9 are moderately invasive. The SCC 12 cell line is characterized by slow growth and minimal invasiveness and has practically no furin expression. (B) Western analysis of PDX expression in transfected cells. The vector alone (Cin)-transfected cells are negative and all PDX-transfected cells show intense expression (PDX). Although SCC 15 and Detroit 562 expressed one form of the protein, A 253 showed two bands corresponding to ≈46 and 50 kDa, that may represent nonglycosylated (46 kDa) and glycosylated (50 kDa) forms. (C) Western analysis of TGF-β1 in cell lysates. Note the relatively higher accumulation of the intracellular pro form (pentagon-shaped arrow) in PDX transfectants (PDX) (compared with the lower expression of Cin vector-alone transfectants). (D) Western analysis of TGF-β1 expression and processing in HNSCC lines transfected with PDX in the conditioned medium. PDX transfectants have reduced levels of the cleavage product (40 kDa, full arrow) and accumulation of the nonprocessed, 50-kDa protein (pentagon-shaped arrow). (E) Western analysis of MT1-MMP showing the processed form (60 kDa, full arrow) in the vector alone-transfected cells (Cin) and the proform (63 kDa, pentagon-shaped arrow) in the PDX-transfected cells (PDX).

Article Snippet: Cell lines derived from human HNSCC (SCC 9, SCC 12, SCC 15, A 253, and Detroit 562) and the human fibrosarcoma cell line HT1080 were obtained from the American Type Culture Collection.

Techniques: Western Blot, Expressing, In Vivo, Transfection, Plasmid Preparation

Journal:

Article Title: Furin inhibition results in absent or decreased invasiveness and tumorigenicity of human cancer cells

doi: 10.1073/pnas.191199198

Figure Lengend Snippet: In vivo invasion assay. (A) Micrograph of a tracheal transplant cross section showing the growth pattern of vector alone-transfected SCC 15 cells. Note that the cells originally placed in the lumen (Lu) have grown inside the tracheal wall and have entered the peritracheal space in the direction of the arrow. Cells invade through the area of least resistance, i.e., the pars membranacea between the opening of the tracheal cartilage ring (Car) as noted by the arrow. (B) Micrograph of a tracheal transplant cross section containing PDX-transfected SCC 15 cells. Note that the cells remain in the luminal (Lu) and interior area of the trachea. (C) Histogram showing the level of penetration of transfected HNSCC cells. Note a marked reduction in the depth of penetration of the PDX-transfected cells into the tracheal wall as compared with their respective vector alone-transfected counterparts (Cin cells). A total of 6–10 tracheal transplants per cell line (total: 24 transplants) were used in two different experiments. One-sided t test indicated statistically significant differences (P < 0.05). (Magnification: A and B, hematoxylin and eosin stain ×30.)

Article Snippet: Cell lines derived from human HNSCC (SCC 9, SCC 12, SCC 15, A 253, and Detroit 562) and the human fibrosarcoma cell line HT1080 were obtained from the American Type Culture Collection.

Techniques: In Vivo, Invasion Assay, Plasmid Preparation, Transfection, H&E Stain