DJ-1 polyclonal antibody (Santa Cruz Biotechnology, Santa Cruz, California, USA) and PTEN monoclonal antibody (Cell Signaling Technology,

Denver). DJ-1 staining was graded according to the intensity and extent of staining of the epithelium as previously described, and HDAC inhibitor immunostaining of all slides were evaluated in a blinded manner [2]. Fluorescent immunohistochemistry To better confirm the cellular location and the relationship between DJ-1 and PTEN in SSCC tissues, fluorescent immunohistochemistry was performed as described previously [27]. Statistical analysis Statistical analysis was performed with the SPSS software (SPSS Standard version 13.0, SPSS). The association of DJ-1 protein expression with SSCC patient’s clinico-pathological Ku-0059436 order features and the recorrelation between molecular features detected with each other were by the χ2 test or Fischer’s exact test. For survival analysis, find more we analyzed all SSCC patients by Kaplan-Meier analysis. Log-rank test was used to compare different survival curves. Multivariate survival analysis was performed on all parameters the Cox regression model. P < 0.05 was considered to be statistically significant. Results DJ-1 and PTEN expression in SSCCs and adjacent non-cancerous tissues DJ-1 was detected mainly in SSCCs and less frequently in adjacent non-cancerous tissues. In comparison, PTEN staining of adjacent non-cancerous tissues

was stronger and more common than that of SSCCs (Figure 1A). To better study the cellular location and the relationship between DJ-1 and PTEN in SSCCs, fluorescent immunohistochemistry was performed, and the results showed that strong expression of DJ-1 isometheptene is found in cytoplasm of SSCC tumor cells, while poor staining of PTEN was observed in cytoplasm of SSCC tumor cells, and that strong expression of PTEN is found in

cytoplasm of adjacent non-cancerous cells, while poor staining of DJ-1 was observed in cytoplasm of adjacent non-cancerous cells (Figure 1B). A summary of DJ-1 and PTEN expression in normal and SSCC tissues is given in Table 2. DJ-1 expression was detected in 88.5% of SSCCs and in 21.0% of adjacent non-cancerous tissues examined, whereas PTEN expression was detected in 46.2% of SSCCs and in 90.5% of adjacent non-cancerous tissues. Moreover, 65.4% of SSCCs were assessed as high grade DJ-1 staining, whereas 78.6% of adjacent non-cancerous tissue had either no or low-grade DJ-1 staining. A significant difference in grade of DJ-1 expression was demonstrated between SSCCs and adjacent non-cancerous tissues (P < 0.001). Further more, we find that DJ-1 expression was linked to lymph nodal status (P = 0.042), pT status (P = 0.037), and UICC stage (P = 0.027), and there was no significant association of overall DJ-1 staining intensity with patient age and tumor grading (Table 3). Figure 1 Expression of DJ-1 in SSCC clinical samples and univariate survival analysis. A.