16 0.36 (0.32–0.40) a. Data are expressed as the mean 2-ΔΔCT (range). Effects of RhoA and RhoC specific shRNA on cell proliferation activity To assess the proliferation activity of tumor cell is important in its invasion and metastasis. Collected

cells were seeded onto 96-well microplates and cellular growths were determined by a continuous 6-day MTT assay. Growth curve was plotted according to these OD value alterations of MTT assay. The difference in cell growth inhibition rate between the HCT116 cells infected with Ad-A1+A2+C1+C2 and the other two groups was not statistically significant in the first 2 days. However, in the third to sixth day, significant differences were found (Fig. 5), but no significant difference between the control cells and the https://www.selleckchem.com/products/poziotinib-hm781-36b.html cells infected with Ad-HK. The results showed that knockdown of RhoA and RhoC see more in the HCT116 cells by shRNA

could change the cell proliferation activity in vitro. Figure 5 displays the growth curve according to the values of 490 nm wavelength light absorption in the three groups. In the third to sixth day, significant difference as exhibited in cell growth inhibition in Ad-A1+A2+C1+C2 group. But there is a slight difference between the control cell and the cells infected with Ad-HK. Invasion and migration power assay in vitro After 22 h incubation, the control HCT116 cells showed stronger invasion activities compared with the ones infected with Ad-A1+A2+C1+C2 group (88 versus 38) (Fig. 6). The differences between

the control and Ad-HK group had no statistical significance. Moreover, the HCT116 cells in Ad-A1+A2+C1+C2 group displayed a significantly lower transmembrane migration activity as compared to those in Ad-HK group and in control HCT116 cells. These findings suggest that RhoA and RhoC expression level seems to be closely associated with the enhanced invasion and migration in HCT116 cell lines. Figure 6 indicates that silencing of RhoA and RhoC may inhibit the invasion and migration of HCT116 cells. The number of invading cells was determined by counting the cells stained with 0.01% crystal violet solution in the lower side of the membrane (A). The graphs (B, C) compare the numbers Florfenicol of transmembrane cells in invasion and migration experiments. Data represent the mean value ± SEM of three independent experiments. *P > 0.05, no significantly difference between the cells treated with Ad-HK and the control cells. **P < 0.05, compared with other groups. Discussion Rho GTPases act as molecular switches to control signal transduction pathways by cycling between a GDP-bound, inactive form and a GTP-bound, active form. Their best-characterized function is in the regulation of actin dynamics. They not only regulate the organization of actin filament system, but also modulate cell motility, proliferation, apoptosis, cell cycle progression, and invasion and metastasis of malignant tumor cells [10, 11].