Inhibition of Melanoma Angiogenesis by Telomere Homolog Oligonucleotides
Figure 3
T-oligo treatment inhibits Matrigel invasion by MM-AN cells and HMVECs. MM-AN cells were treated with 40 M T-oligo or diluent alone. The culture medium was collected after 72 hours. The conditioned medium harvested after 72 hours was used as the chemoattractant for the invasion assay for MM-AN and HMVECs. MM-AN cells were plated on the inserts and allowed 22 hours to move through the pores on the membrane in the bottom of the inserts toward the medium in the lower chamber, interpreted as invasion of the gel. The experimental inserts had a layer of Matrigel, whereas control inserts (not shown) did not. After 22 hours cells that moved through the pores in the membrane were fixed, stained and photographed. (a) Representative images for MM-AN cells are shown. Small open circles are the pores in the membrane, not cells. (b) The total number of cells was counted for 3 membranes for each treatment condition and graphed as a number of cells (mean SEM) for both treatment groups. The assay was repeated twice with identical results. (c) Representative images for HMVECs treated as described for MM-AN above are shown. Small open circles are the pores in the membrane, not cells. (d) The total number of HMVECs were counted for 3 membranes for each treatment condition and graphed as an average number of cells for each treatment group (mean SD). The assay was repeated twice with identical results. Reductions approached but did not reach statistical significance.
