Abstract
BackgroundOur recent study on a panel of human ovarian cancer cells revealed that SKOV-3 cells barely express the Sprouty isoform 1 (Spry1) while 1A9 cells maintain it at a level similar to normal ovarian cells. Here we investigated the functional outcomes of induced alterations in the expression of Spry1 in the two cell lines in vitro.MethodsUsing the Spry1 specific plasmid and siRNA, the expression of Spry1 was induced and conversely silenced in SKOV-3 and 1A9 cells, respectively. The functional outcome was investigated by means of proliferation, MTT, scratch-wound, migration and invasion assays and selection of the stable clones. Mechanism of the effect was explored by Western blot.ResultsIn the Spry1-transfected SKOV-3 cells, a significant reduction in growth and proliferation was evident. Stable clones of the Spry1-transfected SKOV-3 were almost undetectable after day 14. The number of migrated and invaded cells and the percentage of the scratch closure were significantly lower in the Spry1-transfected group. Spry1 silencing in 1A9 cells, on the other hand, led to a significant increase in cell growth and proliferation. The number of migrated and invaded cells and the percentage of the scratch closure significantly increased in Spry1-silenced 1A9 group. Mechanistically, overexpression of Bax, activation of caspases 3, 7, 8 and 9, cleavage of PARP and attenuation of Bcl-2 and Bcl-xl were observed along with reduced activation of Erk and Akt and increased amount and activity of PTEN in the Spry1-transfected SKOV-3 cells.ConclusionsHere, we report the inverse correlation between the expression of Spry1 and growth, proliferation, invasion and migration of ovarian cancer cells.
Highlights
Our recent study on a panel of human ovarian cancer cells revealed that SKOV-3 cells barely express the Sprouty isoform 1 (Spry1) while 1A9 cells maintain it at a level similar to normal ovarian cells
Induced expression of Spry1 is deleterious for viability of the ovarian cancer cell line SKOV-3 We previously demonstrated that the human ovarian cancer cell line SKOV-3 barely expresses Spry1 [8]
Using the trypan blue (Figure 1C) and Methylthiazol tetrazolium (MTT) (Figure 1D) assays, we found that the expression of Spry1 resulted in a significant decrease in the growth and proliferation of the transfected cells at 72 hours post transfection (p-values of 0.0003 and 0.0042 for trypan blue and MTT assay, respectively)
Summary
Our recent study on a panel of human ovarian cancer cells revealed that SKOV-3 cells barely express the Sprouty isoform 1 (Spry1) while 1A9 cells maintain it at a level similar to normal ovarian cells. We investigated the functional outcomes of induced alterations in the expression of Spry in the two cell lines in vitro. The first isoform to be discovered, was initially introduced as a potent feedback inhibitor of the FGF receptor signaling during the tracheal development of the Drosophila embryos [1]. Owing to their regulatory function, deregulation of Sprouty proteins and its contribution. Here we demonstrate how alterations in the protein expression impact functional properties of ovarian cancer cells
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