A Neutral Look At RhoC
The pair matching produced learn more two groups of 40 patients each. The average follow-up period was 88 months (range 2�C120 months). Three separate representative areas of formalin-fixed paraffin embedded (FFPE) tumor tissue were selected for each patient and circled, to match the blocks for the tissue microarray. Each target area on the selected block was punched to form a 0.6-mm diameter tissue core, and these were placed consecutively on recipient blocks of approximately 3?cm?��?2?cm, as previously described [30]. Immunohistochemistry (IHC) was performed on 3?��m sections using a polymer kit purchased from Zytomed Systems (Zytomed Systems Ltd., Berlin, Germany) with one of the following 3 antibodies according to the manufacturer's instructions: E-cadherin (clone CX294, Dako, 1:30 dilution with pretreatment heating in TRS 9 buffer); p53 (clone 8G7G3/2, Zytomed, 1:3000 dilution with pretreatment heating in citric acid buffer) and CDX2 (polyclonal, DakoCytomation, Hamburg, Germany, 1:2500 dilution and no pretreatment). For evaluating the results of the immunohistochemical Z-VAD-FMK stainings, the degree of immunoreactivity of the 3 antibodies (E-cadherin, p53 and CDX2) was quantitatively evaluated. For E-cadherin, a 4-grade scale was used: 0, absence of staining in tumor cells; 1+, weak nuclear and/or cytoplasmic staining in tumor cells; 2+, an intermediate staining intensity between 1+ and 3+ in tumor cells; and 3+, strong nuclear and cytoplasmic staining in tumor cells. For the other markers, a conventional percentage breakdown was used. Thus, a reference range that defines weak and strong expression is given for each marker. For E-cadherin, 0�C1+ signifies weak expression, and all greater values indicate strong expression. For p53, 0�C2% was defined as weak expression, while for CDX2, all values greater than 0�C1% signified strong expression. To guarantee objectivity, two different pathologists carried out the assessment. The statistical analysis was performed using the SPSS statistical software for Windows (version 18.0, SPSS Inc., Chicago, Illinois, USA) and all survival data were analyzed for significance using the log rank test. Comparisons between frequencies were performed using the Chi-square test or, where appropriate, Fisher's exact test. A p-value of RhoC