Increased cell density decreases cysteine proteinase inhibitor activity and increases invasive ability of two prostate tumor cell lines

The ability of a cancer cell to metastasis to a distant site is partly dependent on the secretion of matrix degrading enzymes. The lysosomal cysteine proteinases, cathepsins B and L, have been shown to be secreted by a number of cancer cells and have been implicated in metastasis. Cathepsins B and L are regulated by a class of inhibitors known as the cystatins; aberrant cystatin activity has also been shown in a number of cancer cells. Two prostate cancer cell lines, PC3 and DU145, and a normal prostate epithelial cell (NPC) culture were used to determine the importance of cathepsins L+B and cysteine proteinase inhibitor (CPI) activity in the ability of each cell line to invade the reconstituted basement membrane, Matrigel®. Cathepsin L+B and CPI activities were evident in the cell extract and conditioned media of PC3, DU145 and NPC; however, only the cancer cell lines PC3 and DU145 exhibited invasive ability. Invasive ability was partially inhibited following exposure of PC3 and DU145 cells to the CPI, E-64. Since environmental factors such as cell-cell interactions are responsible for mediating the expression of a number of genes involved in metastasis, the effects of cell density on cathepsin and CPI activities and invasive ability were also determined. CPI activity decreased and invasive ability increased with increasing cell density. We conclude that cathepsin L+B plays a significant role in the invasive ability of the two prostate cancer cell lines, PC3 and DU145. This may be due to decreased regulation by endogenous CPIs whose activity diminishes at high cell densities.