Combination of ouabain and a specific ROCK inhibitor Y27632 in HeLa cells: implication of therapeutic benefits (797.2)

In our previous work we have observed that while the concentrations of ouabain above 100 nM had cytotoxic effects and 30 nM ouabain induced senescence in HeLa cells, ouabain had no noticeable morphological effects on cells at 10 nM and lower concentrations. Therefore in the current work we evaluated the effect of 10nM ouabain on the migration of HeLa cells and the possible involvement of ROCK in this process. The xCELLigence RTCA DP system with CIM‐plates was performed to detect the effect of drugs on cell migration. HeLa cells were exposed to 10 µM Y27632 for 30 min and 0.5 µg/ml CT04 (Rho A inhibitor) for 2h. After preincubation cells were treated with 10 nM ouabain. Cell index was monitored every 15 min for up to 48 h. Our results showed that while 10 nM ouabain had no inhibitory effect on cell migration, combination of ouabain with Y27632 significantly reduced the migrated cell number of HeLa cells. Also Y27632 incubation by itself inhibited the migration of cells. Moreover similar results obtained from CT04 treatment. In contrast with data reported that ouabain has anti migratory properties at nanomolar concentrations, we demonstrated that low nanomolar concentration of the cardiac glycoside did not inhibit the HeLa cell migration. It has been known that ROCK inhibitors have some anticancer properties. In addition, cardiac glycosides have been developed and tested in clinical trial for the treatment of cancer. Therefore our findings imply that combination therapy of ouabain and Y27632 may be beneficial for cancer patients. Grant Funding Source : This study is supported by TUBITAK in 110S426 project.