High Expression of Complement Component 5 (C5) at Tumor Site Associates with Superior Survival in Ewing's Sarcoma Family of Tumour Patients

Abstract

Background. Unlike in most adult-onset cancers, an association between typical paediatric neoplasms and inflammatory triggers is rare. We studied whether immune system-related genes are activated and have prognostic significance in Ewing's sarcoma family of tumors (ESFTs). Method. Data analysis was performed on gene expression profiles of 44 ESFT patients, 11 ESFT cell lines, and 18 normal skeletal muscle samples. Differential expression of 238 inflammation and 299 macrophage-related genes was analysed by t-test, and survival analysis was performed according to gene expression. Results. Inflammatory genes are activated in ESFT patient samples, as 38 of 238 (16%) inflammatory genes were upregulated (P < 0.001) when compared to cell lines. This inflammatory gene activation was characterized by significant enrichment of macrophage-related gene expression with 58 of 299 (19%) of genes upregulated (P < 0.001). High expression of complement component 5 (C5) correlated with better event-free (P = 0.01) and overall survival (P = 0.004) in a dose-dependent manner. C5 and its receptor C5aR1 expression was verified at protein level by immunohistochemistry on an independent ESFT tumour tissue microarray. Conclusion. Immune system-related gene activation is observed in ESFT patient samples, and prognostically significant inflammatory genes (C5, JAK1, and IL8) for ESFT were identified.

SLFN11 Is a Transcriptional Target of EWS-FLI1 and a Determinant of Drug Response in Ewing Sarcoma

“…The gene expression profiles of 44 ES tumor samples and 18 normal skeletal muscle tissues by microarray analysis (26) were downloaded from the Gene Expression Omnibus website (GSE17674, http://www.ncbi.nlm.nih.gov/geo) (27). The raw signal intensities were normalized by Robust Multichip Average (RMA) method and Log 2 -transformed.…”

“…The association between the expression levels of two individual genes in a dataset was evaluated by the Pearson’s correlation, and where multiple comparisons were made, the p-values were adjusted using the Bonferroni method. The Kaplan-Meyer curves based on SLFN11 expression higher or lower than the median in the cohort of 44 ES patients (26) were analyzed using the Mantel-Cox log-rank test. The test results with p < 0.05 were considered to be statistically significant unless otherwise stated.…”

The Ewing Sarcoma Secretome and Its Response to Activation of Wnt/beta-catenin Signaling

“…Pearson correlation coefficients of gene expression and associated confidence intervals in primary patient tumors were determined using three previously reported, independent datasets: GSE 63157 (13), GSE 34620 (24), and GSE 17679 (25). Only data from patient tumors were extracted from the GSE17679 dataset and the average value for duplicate assays from each of 44 unique patient tumors was used.…”

“…Next, we sought to validate whether activation of Wnt/beta-catenin in tumors in vivo is also associated with increased expression of these ECM protein-encoding genes. To achieve this we interrogated three independent, publicly available databases (13,24,25) and assessed correlations between expression of LEF1 and TNC and COL1A1 in primary patient tumors. We have previously shown that LEF1 is a robust biomarker of Wnt/beta-catenin activation in Ewing sarcoma cells and tumors (14).…”

Characterization of Ewing sarcoma associated cancer/testis antigens

“…Expression data were obtained from the Gene Expression Omnibus (GEO) database. 42 The used data sets 19,[43][44][45][46][47][48][49][50][51][52][53][54][55] are summarized in Table 1. Primary data analysis was performed with Expression Console (Affymetrix) applying the Microarray Suite 5 algorithm.…”