Artonin E sensitizes TRAIL-induced apoptosis by DR5 upregulation and cFLIP downregulation in TRAIL-refractory colorectal cancer LoVo cells

Abstract

Background: The TRAIL treatment is an ideal strategy for colorectal cancer (CRC) therapy because of minimal collateral damage to normal cells. Unfortunately, some CRC is TRAIL-refractory cancer, such as LoVo cells. In an effort to overcome TRAIL-refractory cancer, we investigated the effect of artonin E in regulating death receptor 5 (DR5) and cellular FLICE (FADD-like IL-1β-converting enzyme)-inhibitory protein (cFLIP), two major mediators regulate TRAIL-induced apoptosis, in LoVo cells as a model of TRAIL refractory CRC. Methods: TRAIL-refractory cancer (LoVo cells) was treated with artonin E and TRAIL. Cell viability was determined by MTT assay. Apoptotic chromatin condensation was observed by fluorescent Hoechst33342 staining. The mRNA and protein expression of DR5 and FLIP was determined by quantitative PCR and Western blotting analysis, respectively. Results: The combination treatment of artonin E and TRAIL enhanced cytotoxicity and apoptotic chromatin condensation in LoVo cells significantly, while treatment of artonin E or TRAIL alone was not. Artonin E enhanced both mRNA and protein expression of DR5. Interestingly, this is the first report showing that artonin E decreased protein expression of cFLIP. All together we showed that artonin E enhanced TRAIL-induced apoptosis in LoVo cells through DR5 upregulation and cFLIP downregulation. Conclusions: Artonin E was able to increase DR5 expression and decrease cFLIP expression in LoVo cells. These results showed that LoVo cells sensitized TRAIL-induced apoptosis in combined treatment with artonin E and TRAIL. Therefore, the combination treatment of artonin E and TRAIL is one of the potential strategies used for TRAIL-refractory CRC therapy in the future. © Journal of Gastrointestinal Oncology. All rights reserved.