036, 0.076, 0.087, and 0.056 for probes Short,

Probe, Long1, and Long2, respectively) (GSE5206). These data suggest that specific TLR4 transcripts may be incorporated in predictive models of colon cancer survival and recurrence. Lower TLR4 expression and buy ABT-263 microsatellite instability (MSI) Microsatellite unstable tumors are associated with defects in mismatch repair but have improved prognosis [30]. We investigated the relationship between MSI and TLR4 expression. Among 77 microsatellite stable (MSS) and 78 MSI colon cancers, TLR4 expression strongly correlated with MSI status, AZD2014 solubility dmso with MSI tumors having significantly lower TLR4 expression than MSS comparators (GSE13294) [31].MSI was associated with lower expression of TLR4. IHC staining for TLR4 The data from bioinformatics analysis of expression arrays demonstrate an increase in TLR4 genomic expression in neoplastic colon tissue, with relatively selleck high expression in the stromal compartment in particular. We next wished to examine whether this could be seen at the protein level using NCI TMAs. TMAs consisting of 182 independent cancers, 19 adenomas, and matched normal tissue were examined for TLR4 expression (Figure 4A,4B) [11]. Select cores were excluded from the analysis due to poor sample integrity

or staining quality (18 out of 239). The stroma of 82/174 (47.1%) of CRCs was positive for TLR4 expression (score > 3). 62/174 (35.6%) of tumor stroma Fludarabine mw were strongly positive (score > 5). The epithelium of 11/174 (6.32%) of CRCs was positive for TLR4 expression; 6/174 (3.45%) of tumor epithelia were strongly positive. Figure 4 Immunofluorescent staining of TMAs. A) Low power (10x) view of NCI TMA slide stained for TLR4 (green), intestinal epithelium/pan-cytokeratin (red), and nucleus/DAPI (blue). B) Representative tissue cores from normal (I), adenomatous polyps (II), and CRC (III and IV) are shown. C and D)

TLR4 staining score by tissue type and tissue compartment (stroma vs epithelium) are shown. C) TLR4 staining in the tumor stroma had a significantly higher average intensity score for stages 3 and 4 CRC when compared to stage 1. D) TLR4 staining in the tumor epithelium had a significantly higher average intensity score for stages 2 and 3 when compared to stage 1. E) TLR4 staining by compartment broken down by stage (controlling for grade) and grade (controlling for stage). TLR4 staining in the tumor stroma and epithelium increases with tumor stage Using semi-quantitative scoring, a positive relationship was noted between TLR4 staining score in the tumor stroma and tumor stage, controlling for histology grade, with significantly higher intensity score for stages 3 and 4 compared to stage 1 (Stage 1 = 2.80, Stage 2 = 3.24, Stage 3 = 4.36, Stage 4 = 3.75; p = NS, 0.0004, and 0.04, respectively) (Figure 4C,4D,4E).