80% to 23.74%, and the healing rates at 12 h, 24 h and 36 h (p < 0.001). By selleck compound contrast, the healing rate of NPC 5-8 F cells was not affected by treatment of lipofectamine alone and transfection of pEGFP-C3 and PinX1-FAM-siRNA (p > 0.05). Figure 6 Effect of PinX1 on wound healing ability of nasopharyngeal carcinoma

5-8 F cells in scratch assay. Cells transfected with pEGFP-C3-PinX1 (a), pEGFP-C3 (b) and PinX1-FAM-siRNA(e), treated with lipofactamine alone (c), and untreated (d) were inoculated in 6-well plates pre-coated with collagen IV, cultured in media containing 10% newborn calf serum till forming monolayer, then scratched and photographed at 0 h, 12 h, 24 h and 36 h after scratching. The results show that overexpression of PinX1 by transfection of pEGFP-C3-PinX1 significantly increased the wound healing time see more of NPC 5-8 F cells, while downregulation of PinX1 by transfection of FAM-siRNA reduced has no effect on wound healing. We then examined the effect of PinX1 on hTERT mRNA level and telomerase activity. As shown in Tables 4 and 5 and Figures 7 and 8, overexpression of Pin X1 by transfection of pEGFP-C3-PinX1 significantly reduced hTERT mRNA level by 21% and decreased

telomerase activity in NPC 5-8 F cells (p = 0.000). By contrast, reduced PinX1 by transfection of PinX1-FAM-siRNA had effects on neither hTERT mRNA eltoprazine level nor telomerase activity in NPC 5-8 F cells (p > 0.05). In addition, hTERT mRNA level and telomerase activity in NPC 5-8 F cells were not affected by transfection of pEGFP-C3 and treatment of lipofectamine alone. Table 4 hTERT

mRNA level in each group Sample hTERT mRNA F P pEGFP-C3-PinX1 0.789 ± 0.024* 117.689 0.000 pEGFP-C3 0.978 ± 0.011     Lipofectamine alone 0.987 ± 0.014     Untreated 1.000 ± 0.000     PinX1-FAM-siRNA 1.001 ± 0.085**     * vs untreated, P < 0.001, ** vs untreated, P > 0.05. hTERT mRNA level was normalized to GAPDH. Table 5 Telomerase activity in NPC cells Samples Telomerase activity F P pEGFP-C3-PinX1 36227.63 ± 2181.748* 53.816 0.000 pEGFP-C3 58346.993 ± 2181.748     Lipofectamine alone 59697.199 ± 2181.748     Untreated 62552.354 ± 2181.748     PinX1-FAM-siRNA 63600.608 ± 2181.748**     * vs untreated, P < 0.001; ** vs untreated, P > 0.05. Figure 7 Effects of PinX1 on hTERT mRNA level in NPC 5-8 F cells. PinX1 mRNA levels in NPC 5-8 F cells transfected with (a) pEGFP-C3-PinX1, (b) with pEGFP-C3, (c) treated with lipofectamine alone, (d) untreated and (e) transfected with PinX1-FAM-siRNA were measured in RT-PCR and normalized to internal control GAPDH. Data were presented as mean value of three experiments showing that overexpression of PinX1 significantly decreased hTERT mRNA level. Figure 8 Effect of PinX1 on telomerase activity in nasopharyngeal carcinoma cells.