The results showed that the level of LATS1 expression was an independent prognostic factor for glioma (P<0.001) (Table 3). Figure 2 Reexpression of LATS1 in glioma U251 cells. A. Real-time PCR analysis indicated the highest mRNA expression of LATS1 in two cell clones pLATS1-2 and −4. B. Western blotting assay shows significantly increased protein expression of LATS1 in pLATS1-2 and −4 suppressed the expression of cell cycle factor CCNA1 protein compared to Control-vector

cells. β-actin was used as the internal control. Table 3 Summary of univariate and multivariate Cox regression analysis of overall survival duration Parameter Univariate analysis Multivariate analysis P HR 95%CI P HR 95%CI Age ≥55vs. <55 years 0.069 0.777 0.593-1.019       Gender Male vs. female 0.160 0.820 0.621-1.082       WHO grade Ivs.II vs.III vs.IV 0.000 1.715 1.454-2.023

0.000 1.463 1.233-1.735 KPS ≥80 vs. < 80 0.000 2.033 1.540-2.684 this website 0.000 2.437 1.810-3.283 LAST1 expression             Strong vs.Positive vs.Weak vs.Negative* 0.000 0.437 0.362-0.528 0.000 0.389 0.316-0.478 Overexpression of LATS1 in glioma U251 cells To study its biological functions, we introduced the LATS1 gene into the glioma U251 cell line using pCDF-GFP lentivirus expression vector. Five (5) stably transfected cell clones were obtained. Real-time PCR identified two cell clones (LATS1-2,-4) with the highest mRNA expression of LATS1 (Figure 2A). Further, LATS1 protein was highly expressed in two cell clones by western blotting assay with LATS1 antibody,while control clone cells lacked similar expression (Figure 2B). RAD001 research buy LATS1 inhibits cell proliferation in vitro To analyze the function of LATS1, we studied the rate of cell proliferation of LATS1-expressing LATS1-2 and −4 cells. The growth curves determined by MTT assay revealed that LATS1 significantly inhibited

cell proliferation of these two lines of cells compared to control clone cells (Figure 3A). In a colony formation assay LATS1-overexpressing LATS1-2 and −4 cells formed significantly less colonies than control clone cells (P < 0.001 ASK1 for both cell types) (Figure 3B, Table 4), suggesting the inhibitory effect of LATS1 on anchorage-dependent growth of glioma cells. Figure 3 Overexpression of LATS1 inhibted cell proliferation in vitro. A. The cell growth of Control-vector cells and pLATS1-2 and −4 cells, were examined by MTT assay over a seven-day period. *P < 0.05, as compared to control-vector cells. B. The cell growth of control-vector cells and pLATS1-2 and −4 cells, were examined by plate colony formation assay. *P < 0.05, as compared to control-vector cells. Table 4 Plate clone formation assay among pLATS1-2, pLATS1-4, and Ctr-vector cells Cells Number P value pLATS1-2 45.33 ± 4.16   pLATS1-4 34.67 ± 6.25   Ctr-vector 77.33 ± 7.12 p<0.