Each sample was examined in triplicate and the amounts of the PCR products produced were non-neoplasticized to GAPDH which served as internal

control. Statistical analysis All computations were carried out using the software of SPSS version13.0 for Windows (SPSS Inc, IL, USA). Data were expressed as means±standard deviation (SD). The analysis of variance (ANOVA) was used to determine the statistical differences among the groups. A life table was selleck inhibitor calculated according to the Kaplan-Meier method. Hazard ratios for the time-to-event endpoint were estimated using the multivariate Cox regression analysis in a forward stepwise method to evaluate the effect of multiple independent prognostic factors on survival outcome. Differences were considered statistically significant when p was less than 0.05. Results CLIC1 mRNA expression in human glioma tissues Selleck NVP-LDE225 The expression levels of CLIC1 mRNA were detected in 20 glioma and 10

non-neoplastic brain tissues normalized to GAPDH. As shown in Figure 1A, the expression levels of CLIC1 mRNA were found to be distinctly increased in glioma tissues compared to non-neoplastic brain tissues, corresponding to the glioma WHO grades. The statistic results (Figure 1B) showed that its expression in high-grade (III-IV; 2.2±0.08) and low-grade (I-II; 1.6±0.06) gliomas were both significantly higher than that in non-neoplastic brains tissues (0.3±0.01; Acyl CoA dehydrogenase both P<0.001). Additionally, there was also a significant difference in mRNA copies of CLIC1 between high-grade (III-IV) and low -grade (I-II) glioma tissue specimens (P=0.002). Figure 1 CLIC1 mRNA expression in 20 glioma tissues with different grades and in non-neoplastic brain tissues were detected by real-time quantitative RT-PCR assay. (A) Expression levels of CLIC1 mRNA in glioma tissues with different grades and non-neoplastic brain tissues. (B) A graphical representation

of the CLIC1 mRNA level expression profiles in (A). ‘N’ refers to non-neoplastic brain tissues; ‘I~II’ refers to glioma tissues with grade I~II; ‘III~IV’ refers to glioma tissues with grade III~IV. Elevated expression of CLIC1 protein in human glioma tissues The expression of CLIC1 protein was detected in 128 glioma specimens and 10 nonneoplastic brain tissues using immunohistochemical staining. Representative photographs of CLIC1 immunostainings were shown in Figure 2. In the glioma sections, CLIC1 was mainly detected in the cytoplasm (Figure 2A), which was consistent with previous studies on other cancers [10–12]. In contrast, the non-neoplastic brain tissues expressed a trace amount of CLIC1 (Figure 2B). CLIC1 was not present in negative controls with non-immune IgG (Figure 2C) and in normal gastric tissues (Figure 2D).