TRAIL mutant membrane penetrating peptide alike-MuR6-TR enhances the antitumor effects of TRAIL in pancreatic carcinoma both in vitro and in vivo

摘要

To remedy the drug resistance of natural tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and enhance its antitumor effects, we prepared a type of TRAIL mutant membrane penetrating peptide alike (TMPPA)?TRAIL mutant?R6 (MuR6-TR) by mutating the N?terminal of the soluble TRAIL gene sequence. The expressed MuR6?TR protein was purified to treat pancreatic carcinoma cell lines BxPC?3 and PANC?1. The inhibitory effects on the proliferation of BxPC?3 and PANC?1 cells was assessed with CCK?8 assay and compared with natural TRAIL. The antitumor effect of MuR6?TR was assessed on implant tumors derived from PANC?1 cells in nude mice and compared with gemcitabine. Finally, the soluble MuR6?TR gene was successfully mutated with 4 amino acids in the N?terminal of TRAIL and had a molecular size of 513?bp. The mutant MuR6?TR was connected to pET32a and verified by enzymatic digestion and sequencing. The recombinant MuR6?TR was transformed and expressed in Escherichia?coli. The CCK?8 assay results indicated that MuR6?TR inhibited the growth of BxPC?3 and PANC?1 cells in a dose?dependent manner, with IC50 values of 4.63?and?7.84?ng/ml, respectively, which were much lower than that of natural TRAIL. MuR6?TR demonstrated a higher inhibitory effect on tumor growth (24.2%) than natural TRAIL (14.4%) and an effect similar to that of gemcitabine at an early period. Thus, the mutant MuR6?TR exhibited a stronger antitumor effect than that of natural TRAIL both in?vivo and in?vitro and may have potential therapeutic value for pancreatic carcinoma, which requires further validation.