Inflammatory Protein Serum Amyloid A1 Marks a Subset of Conventional Renal Cell Carcinomas with Fatal Outcome  Claudia Paret, Zorica Schön, Adrianna Szponar, Gyula Kovacs  European Urology  Volume 57, Issue 5, Pages 859-866 (May 2010) DOI: 10.1016/j.eururo.2009.08.014 Copyright © 2009 Terms and Conditions

Fig. 1 Overexpression of SAA1 in aggressive conventional RCC. (A). Selection of the 50 best fitting genes by GSEA which are overexpressed in aggressive conventional RCCs. Blue colour indicates low expression, red colour indicates high expression. Each row in the heat map represents a gene, and each column represents a tumour sample. (B). Quantitative RT-PCR was performed with 20 non-aggressive conventional RCC samples (grey columns) and 24 aggressive conventional RCC (black columns). The identification number of the tumours is shown. Expression level was normalised to actin and expression level more than 200 was considered positive. European Urology 2010 57, 859-866DOI: (10.1016/j.eururo.2009.08.014) Copyright © 2009 Terms and Conditions

Fig. 2 Expression of SAA1 protein in conventional RCC tissues. A. Negative SAA1 staining of a non-aggressive conventional RCC (×200). B. Moderate SAA1 staining on the plasma membrane of cancer cells (×200). C. A variable positive staining of cytoplasm of tumour cells (×200). D. In addition to the cytoplasmic staining small SAA1 globlets were seen in some of the cells (×630, enlargement of C). European Urology 2010 57, 859-866DOI: (10.1016/j.eururo.2009.08.014) Copyright © 2009 Terms and Conditions

Fig. 3 Correlation between SAA1 protein expression and patient’s survival. Kaplan-Meier survival analysis was performed with 72 patients. N = negative SAA1 staining, P = positive SAA1 staining. European Urology 2010 57, 859-866DOI: (10.1016/j.eururo.2009.08.014) Copyright © 2009 Terms and Conditions

Fig. 4 SAA1 stimulated MMP-9 expression and invasion potential of conventional RCC cells. (A). SAA1 stimulates MMP-9 expression at the protein level in cell lines HD2, HA344 and HA465. The supernatant of the cell lines without and with stimulation by adding SAA1 of 10μg/ml was collected and analysed by zymography. (B). The conventional RCC cell line HA344 stimulated with increasing concentrations of SAA1 showed an increasing activity of MMP-9 but not of MMP-2 by zymography. (C). The conventional RCC cell line HA344 was pretreated with PTX and SAA1 alone or in combination. Notice that SAA1 induced expression of MMP-9 was not suppressed by adding PTX. (D). SAA1 stimulates MMP-9 expression at the mRNA level in cell lines HD2, HA344 and HA465, which corresponds to the expression at the protein level as shown under A. (E). SAA1 stimulates the invasiveness of conventional cell line HD2 (p<0.05) but not of the conventional cell line HD48 (p=0.4). The number of migrating cells of cell line HD2 was at least three times higher after SAA1 treatment. European Urology 2010 57, 859-866DOI: (10.1016/j.eururo.2009.08.014) Copyright © 2009 Terms and Conditions