Effects of Recombinant Circularly Permuted Tumor Necrosis Factor (TNF)-Related Apoptosis-Inducing Ligand (TRAIL) (Recombinant Mutant Human TRAIL) in Combination with 5-Fluorouracil in Human Colorectal Cancer Cell Lines HCT116 and SW480

Abstract

BackgroundCircularly permuted tumor necrosis factor-related apoptosis-inducing ligand, a mutant form of tumor necrosis factor-related apoptosis-inducing ligand, is an effective antitumor cytokine. However, its antitumor effect in colorectal cancer is unclear. This study assessed the antitumor effect of circularly permuted tumor necrosis factor-related apoptosis-inducing ligand alone or with 5-fluorouracil in colorectal cancer cells in vitro and explored the underlying mechanisms.Material/MethodsWe used the (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) assay to analyze cell proliferation inhibition. The apoptotic effects of circularly permuted tumor necrosis factor-related apoptosis-inducing ligand, 5-fluorouracil, or both in human colorectal cancer cells were evaluated using flow cytometry. Furthermore, the levels of apoptosis-related proteins were examined by Western blotting.ResultsCompared to either agent alone, cotreatment with 5-fluorouracil and circularly permuted tumor necrosis factor-related apoptosis-inducing ligand showed obvious antitumor effects and induced significant apoptosis of colorectal cancer cells. 5-Fluorouracil enhanced circularly permuted tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis by increasing death receptor 4 and 5 levels in HCT116 cells, but only of death receptor 4 in SW480 cells. Moreover, 5-fluorouracil plus circularly permuted tumor necrosis factor-related apoptosis-inducing ligand increased apoptosis-related protein levels such as cleaved caspase-3, caspase-8, and poly-ADP-ribose polymerase and downregulated that of the survival protein B-cell lymphoma-extra-large. Pretreatment with the pan-caspase inhibitor, z-VAD-FMK, attenuated the caspase-dependent apoptosis induced by circularly permuted tumor necrosis factor-related apoptosis-inducing ligand alone or combined with 5-fluorouracil.ConclusionsCotreatment with 5-fluorouracil and circularly permuted tumor necrosis factor-related apoptosis-inducing ligand showed enhanced antitumor effects on colorectal cancer cells.

Research Progress on the Relationship Between Inflammation and Colorectal Cancer

“…74,75 TNFα can induce the expression of genes involved in tumor invasion and metastasis by activating NF-κB, including adhesion molecules, matrix metallopeptidase 9 (MMP9), cyclooxygenase-2 (COX-2), and vascular endothelial growth factor (VEGF). 75,76 In addition, TNFα can also trigger the activation of the epithelial-mesenchymal transition (EMT). 77 These characteristics can promote the formation and metastasis of CRC neovascularization.…”

“…Tumor necrosis factor‐α (TNF‐α) can induce tumor cell apoptosis under normal circumstances, but under pathological conditions it acts as an inflammatory factor to promote tumor development 74,75 . TNF‐α can induce the expression of genes involved in tumor invasion and metastasis by activating NF‐κB, including adhesion molecules, matrix metallopeptidase 9 (MMP9), cyclooxygenase‐2 (COX‐2), and vascular endothelial growth factor (VEGF) 75,76 . In addition, TNF‐α can also trigger the activation of the epithelial–mesenchymal transition (EMT) 77 .…”

5-Fluorouracil enhances the chemosensitivity of gastric cancer to TRAIL via inhibition of the MAPK pathway

“…5-FU and its derivatives, such as capecitabine, tegio, carmo uor, etc., are currently the most common drugs used in rst-line chemotherapy for gastric cancer (7), and these drugs prevent tumors by inhibiting the activity of thymidylate synthase and the production of DNA in cells (8). In some cancer types, researchers have explored the use of 5-FU as a TRAIL sensitizer to synergistically exert an antitumor effect (9)(10)(11). However, the mechanism by which 5-FU induces gastric cancer cells sensitivity to TRAILinduced apoptosis has not been reported.…”

Mechanisms underlying synergism between circularized tumor necrosis factor‐related apoptosis inducing ligand and bortezomib in bortezomib‐sensitive or ‐resistant myeloma cells

“…This led to the development of circularly permutated TRAIL (CPT), a cyclization allosteric form of wild‐type TRAI 13 . Compared to wild‐type TRAIL, CPT has greater antitumor activity, and better stability/biological activity in aqueous solution 14 . In a phase II trial of CPT monotherapy, among 27 MM patients with relapsed/refractory multiple myeloma (RRMM), 1 patient achieved a near‐complete response, and 8 patients achieved partial responses 15 .…”