Purpose: We previously found that cellular FLICE-inhibitory protein (c-FLIP), caspase 8, and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor 2 (DR5) are major regulators of cell viability and chemotherapy-induced apoptosis in colorectal cancer. In this study, we determined the prognostic significance of c-FLIP, caspase 8, TRAIL and DR5 expression in tissues from patients with stage II and III colorectal cancer.Experimental Design: Tissue microarrays were constructed from matched normal and tumor tissue derived from patients (n = 253) enrolled in a phase III trial of adjuvant 5-fluorouracil-based chemotherapy versus postoperative observation alone. TRAIL, DR5, caspase 8, and c-FLIP expression levels were determined by immunohistochemistry.Results: Colorectal tumors displayed significantly higher expression levels of c-FLIP (P < 0.001), caspase 8 (P = 0.01), and DR5 (P < 0.001), but lower levels of TRAIL (P < 0.001) compared with matched normal tissue. In univariate analysis, higher TRAIL expression in the tumor was associated with worse overall survival (P = 0.026), with a trend to decreased relapse-free survival (RFS; P = 0.06), and higher tumor c-FLIP expression was associated with a significantly decreased RFS (P = 0.015). Using multivariate predictive modeling for RFS in all patients and including all biomarkers, age, treatment, and stage, we found that the model was significant when the mean tumor c-FLIP expression score and disease stage were included (P < 0.001). As regards overall survival, the overall model was predictive when both TRAIL expression and disease stage were included (P < 0.001).Conclusions: High c-FLIP and TRAIL expression may be independent adverse prognostic markers in stage II and III colorectal cancer and might identify patients most at risk of relapse. Clin Cancer Res; 16(13); 3442-51. ©2010 AACR.Colorectal cancer is the second most common cause of cancer-related deaths. It is now well established that in patients with stage III disease undergoing curative surgical resection, adjuvant 5-fluorouracil (5-FU)-based chemotherapy reduces tumor recurrence rates and improves overall survival (1-3). However, as many as 65% of patients with stage III colorectal cancer are cured by surgery alone (4). In stage II disease, the QUASAR trial, comparing adjuvant chemotherapy versus observation alone, concluded that although some patients benefited from adjuvant therapy, the improvement in 5-year survival was small (3.6%), and >80% of stage II patients were cured by surgery alone (2). Thus, the identification and validation of prognostic biomarkers of relapse in stage II and III colorectal cancer are urgently needed to spare patients, who could be cured by surgery alone, from unnecessary treatment with chemotherapy. This has become even more important following the addition of oxaliplatin to a fluoropyrimidine as an option in adjuvant therapy, as there is now a higher potential for longer term toxicity such as sensory neuropathy.The aim of this study is to use immunohi...

We have isolated several lines of rat embryo cells transformed by simian virus 40. All these lines are fully transformed with regard to saturation density and serum sensitivity, but they differ greatly in their anchorage dependence, as assayed by efficiency of plating in methyl cellulose suspension. This set of lines reveals a consistent relation of plasminogen activator production to plating efficiency in methyl cellulose. T-antigen-positive transformed lines that synthesize activator grow in methyl cellulose suspension, while T-antigen-positive transformed lines that do not synthesize activator fail to form colonies in suspension. Normal rat embryo cells produce very little plasminogen activator and do not grow in methyl cellulose.Sera that permit high levels of plasmin formation and activity support growth in semi-solid medium better than sera whose plasminogen is activated poorly and/or sera that contain inhibitors to plasmin.When a small DNA tumor virus, such as simian virus 40 (SV40) or polyoma, infects susceptible mammalian fibroblasts in vitro, a number of physiological and social characteristics of the cells are drastically and permanently altered. The sum of these behavioral changes, especially the loss of sensitivity to the growth-inhibiting effects of suspension in semi-solid media, extensive cell-cell contact, and serum-depletion, is termed transformation (1-8). The multiplicity of changes has been a major problem in understanding the relation of viral transformation to in vivo tumorigenicity (9-15). However, loss of sensitivity to these three different environmental stimuli is not coordinate, and transformants have been isolated that are still sensitive to one or even two of them (4,7,8).The demonstration that analysis of nonselected SV40 transformation permits easy isolation of clonal transformed lines having widely differing in vitro properties (7) encouraged us to attempt to determine whether plasminogen activator production, a biochemical alteration which may be related to malignancy (16-27), was correlated with any of the above mentioned measures of transformation. In this paper we report the correlation of anchorage dependence with production of plasminogen activator in a set of SV4O-transformed clones derived from infected rat embryo cells. A preliminary report of this work has appeared elsewhere (28). METHODS

Background:The CXC-chemokine expression is linked with colorectal cancer (CRC) progression but their significance in resected CRC is unclear. We explored the prognostic impact of such expression in stage II and III CRC.Methods:Tissue microarrays were constructed from stage II and III CRC biopsies (n=254), and the expression of CXCL1 and CXCL8, and their receptors CXCR1 and CXCR2, in malignant and adjacent normal tissue was graded by immunohistochemistry and was correlated with prognostic factors.Results:Expression of CXCL1, CXCR1 and CXCR2 was elevated in tumour epithelium relative to normal adjacent tissue (P<0.001). CXCL8 expression was detectable in the peritumoural inflammatory infiltrate. There was no overall association between CXCL1, CXCR1 or CXCR2 expression and prognostic endpoints; however, univariate subgroup survival analysis demonstrated an inverse association between CXCL1 and recurrence-free survival (RFS) in stage III patients (P=0.041). The CXCL8 positivity in the tumour infiltrate, however, correlated with earlier disease stage (P<0.001) and improved relapse-free survival across the cohort (P<0.001). Disease stage (P<0.001) and tumour infiltrate CXCL8 positivity (P=0.007) were associated with enhanced RFS in multivariate Cox regression analysis.Conclusion:Autocrine CXC-chemokine signalling may have adverse prognostic effects in early CRC. Conversely, CXCL8 positivity within the immune infiltrate may have good prognostic significance.