DR5 and DcR2 are expressed in human lumbar intervertebral discs.

STUDY DESIGN: To conform the 3 media way of the apoptosis for the degenerative lumbar disc with DR5 (TRAIL-R2) and DcR2 (TRAIL-R4), as one of the tumor necrosis factors family. OBJECTIVE: To detect the expression of the DR5 (TRAIL-R2) and DcR2 (TRAIL-R4) protein and mRNA in human herniated and normal lumbar intervertebral discs (IVD). SUMMARY OF BACKGROUND DATA: The pathogenesis of lumber intervertebral disc herniation and degeneration is still unclear. A series of reports have suggested that apoptosis may play a key role in intervertebral disc degeneration. There are 3 apoptosis-inducing factors: FasL, TNF-alpha, and TRAIL, which trigger cell death by apoptosis-signaling pathways. These factors combined with the ligand to induce apoptosis. We have reported the expression of DR4 in IVD before. To our knowledge, there are still no studies the important role of the DR5 and DcR2 for apoptosis in IVD tissue. METHODS: The expression and distribution of DR5 and DcR2 proteins were assessed using immunostaining in 60 herniated lumbar IVD and 22 normal lumbar IVD tissue samples. DR5 and DcR2 mRNA was also quantified using real time fluorescent reverse transcriptase-polymerase chain reaction (RT-PCR) in 30 herniated lumbar IVD and 9 normal lumbar IVD. RESULTS: There were significant differences in the percentage of samples with DR5 expression between the herniated (41.60%) and normal IVD (26.09%) groups (P = 0.001). Similarly, DR5 mRNA levels differed between groups (P = 0.025). However, there were no differences in DcR2 protein or mRNA levels. CONCLUSION: The current results indicate that disc cells, after herniation, undergo apoptotic cell death via the DR5/TRAIL pathway.

High TRAIL death receptor 4 and decoy receptor 2 expression correlates with significant cell death in pancreatic ductal adenocarcinoma patients.

OBJECTIVES: The importance of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and TRAIL receptor expression in pancreatic carcinoma development is not known. To reveal the putative connection of TRAIL and TRAIL receptor expression profile to this process, we analyzed and compared the expression profile of TRAIL and its receptors in pancreatic tissues of both noncancer patients and patients with pancreatic ductal adenocarcinoma (PDAC). METHODS: Thirty-one noncancer patients and 34 PDAC patients were included in the study. TRAIL and TRAIL receptor expression profiles were determined by immunohistochemistry. Annexin V binding revealed the apoptotic index in pancreas. Lastly, the tumor grade, tumor stage, tumor diameter, perineural invasion, and number of lymph node metastasis were used for comparison purposes. RESULTS: TRAIL decoy receptor 2 (DcR2) and death receptor 4 expression were up-regulated in PDAC patients compared with noncancer patients, and the ductal cells of PDAC patients displayed significant levels of apoptosis. In addition, acinar cells from PDAC patients had higher DcR2 expression but lower death receptor 4 expression. Increased DcR2 expression was also observed in Langerhans islets of PDAC patients. CONCLUSIONS: Differential alteration of TRAIL and TRAIL receptor expression profiles in PDAC patients suggest that the TRAIL/TRAIL receptor system may play a pivotal role during pancreatic carcinoma development.

TNF-alpha and shear stress-induced large artery adaptations.

BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) up-regulation has been associated with both low and high shear-induced arterial remodeling. To address this apparent paradox and to define the biology of TNF-alpha signaling in large arteries, we tested the hypotheses that differential temporal expression of TNF-alpha drives shear-regulated arterial remodeling. MATERIALS AND METHODS: Both low- and high-shear environments in the same rabbit were surgically created for common carotid arteries. Common carotid arteries (n = 60 total) were harvested after d0, d1, d3, d7, and d14 and analyses included morphology, TNF-alpha, and IL-10 mRNA quantitation. In separate experiments, animals received pegylated soluble TNF-alpha Type 1 receptor (PEG sTNF-RI) or vehicle via either short- or long-term dosing to define the effect of TNF-alpha blockade. RESULTS: The model yielded a 14-fold shear differential (P < 0.001) with medial thickening under low shear (P = 0.025), and evidence of outward remodeling with high shear (P = 0.007). Low shear immediately up-regulated TNF-alpha expression approximately 50 fold (P < 0.001) at d1. Conversely, high shear-induced delayed and sustained TNF-alpha expression (22-fold at d7, P = 0.012; 23-fold at d14, P = 0.007). Both low and high shear gradually induced IL-10 expression (P = 0.002 and P = 0.004, respectively). Neither short-term (5-day) nor long-term (14-day) blockage of TNF-alpha signaling resulted in treatment-induced changes in the remodeling of low- or high-shear arteries. CONCLUSIONS: Shear stress differentially and temporally regulates TNF-alpha expression in remodeling large arteries. However, TNF-alpha blockage did not substantially impact the final shear-induced morphology, suggesting that large arteries can remodel in response to flow perturbations independent of TNF-alpha signaling.

Membrane expression of DR4, DR5 and caspase-8 levels, but not Mcl-1, determine sensitivity of human myeloma cells to Apo2L/TRAIL.

The improved recombinant form of the death ligand Apo2L/TRAIL (Apo2L/TRAIL.0) is not cytotoxic for normal human cells and is a good candidate for the therapy of multiple myeloma (MM), a B-cell neoplasia that remains incurable. We have analyzed the molecular determinants of myeloma sensitivity to Apo2L/TRAIL.0 in a number of MM cell lines, the mechanisms of resistance and a possible way of overcoming it. Expression of one death receptor for Apo2L/TRAIL (DR4 or DR5) is sufficient to transduce death signals, though DR5 was more efficient when both receptors were present. Membrane expression of decoy receptors (DcR1, DcR2) and intracellular levels of c-FLIP(L), XIAP and Mcl-1 were not predictive of resistance to Apo2L/TRAIL. Inhibition of Mcl-1 degradation did not prevent Apo2L/TRAIL-induced apoptosis. In IM-9 cells, resistance was associated to a reduced caspase-8 expression. U266 cells, though expressing significant levels of DR4 and caspase-8, were nevertheless resistant to Apo2L/TRAIL. This resistance could be overcome by co-treatment with valproic acid (VPA), a histone deacetylase inhibitor. VPA caused the redistribution of DR4 to plasma membrane lipid rafts and restored DR4 signaling. Overexpression of Mcl-1 in U266 cells did not prevent Apo2L/TRAIL cytotoxicity in VPA-sensitized cells. These results, taken together, support the possible use of Apo2L/TRAIL.0 in the treatment of MM.