The inva sion chambers were then incubated at 37 C for 24 h. After incubation, the inserts and cells on the upper side of the filter were removed. The filters were fixed, mounted, and stained according to the manufacturers instructions. The cells that invaded to the underside of the filter were counted. Each experiment was repeated selleck chemicals FTY720 three times. Inhibitors,Modulators,Libraries The values obtained were calculated by averaging the total number of cells from triplicate determinations. Statistical analysis Statistical analysis was performed using SPSS 17. 0. All data were expressed as meansstandard error of the mean. The comparisons between mRNA and pro tein expression levels in one group between tumour and matched clear surgical margin tissues and in the other group between tumour and metastatic lymph nodes were made by the paired sample t test for parametric analysis or Wilcoxon signed rank test for nonparametric analysis.

Comparisons related to age or sex in clinical characteris tics were made by the Mann Whiney U test. The differ Inhibitors,Modulators,Libraries ences between experimental and control Inhibitors,Modulators,Libraries groups and among gene expression levels related to clinical stages were analysed by one way analysis of variance. The correlation Inhibitors,Modulators,Libraries between mRNA and protein levels was analysed by Spearman rank correlation. Statistical signifi cance was assumed for a two tailed p 0. 05. Results Reduced expression of 14 3 3epsilon in LSCC 14 3 3epsilon mRNA levels in 72 of the 101 cases of LSCC and matched clear surgical margin tissues were evaluated by RT PCR. The result showed that the 14 3 3epsilon mRNA expression level in LSCC tissues was sig nificantly lower than that in clear surgical margin tissues.

14 3 3epsilon protein levels were detected in all 101 cases of LSCC tissues and matched clear surgical margin tissues and in all 9 cases of metastatic lymph node tissues by western blotting. The 14 3 3epsilon protein expression Inhibitors,Modulators,Libraries level in LSCC tissues was significantly lower than that in clear surgical margin tissues. However, there was no significant correlation between mRNA levels and protein levels. The 14 3 3epsilon pro tein expression scientific research level in the metastatic lymph nodes was also lower than that in matched LSCC tissues. We assessed the 14 3 3epsilon expression levels with respect to clinical characteristics. No differences were identified in protein and mRNA levels of 14 3 3epsilon with respect to patient age and sex. There was no difference in mRNA levels with respect to patient clinical stage. However, the protein expression level of 14 3 3epsilon in stage III or IV tumours was significantly lower than that in any stage I or II tumours. Decreased proliferation of Hep 2 cells transfected with 14 3 3epsilon Hep 2 cells were transfected with 14 3 3epsilon GFP and GFP expression vectors.