TRAIL mutant membrane penetrating peptide alike-MuR6-TR enhances the antitumor effects of TRAIL in pancreatic carcinoma both in vitro and in vivo.
TRAIL mutant membrane penetrating peptide alike-MuR6-TR enhances the antitumor effects of TRAIL in pancreatic carcinoma both in vitro and in vivo.
复制标题
TRAIL突变型穿膜肽类似物-MuR6-TR增强TRAIL在胰腺癌体内和体外的抗肿瘤作用
DOI：
10.3892/ijmm.2017.2968
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发表时间：
2017-06
影响因子：
5.4
通讯作者：
Yi C
中科院分区：
文献类型：
--
作者：
Sun L;Chen C;Zhu A;Huang Y;Zhu H;Yi C
为了弥补天然肿瘤坏死因子相关凋亡诱导配体（TRAIL）的耐药性，增强其抗肿瘤作用，我们通过突变可溶性TRAIL基因序列的n端，制备了一类TRAIL突变体穿膜肽样物(TMPPA)-TRAIL突变体R6 （MuR6-TR）。纯化表达的MuR6-TR蛋白用于治疗胰腺癌细胞株BxPC-3和PANC-1。CCK-8法评价其对BxPC-3和PANC-1细胞的增殖抑制作用，并与天然TRAIL进行比较。评估MuR6-TR对裸鼠PANC-1细胞源性植入瘤的抗肿瘤作用，并与吉西他滨进行比较。最后，成功突变了TRAIL n端4个氨基酸的可溶性MuR6-TR基因，其分子量为513 bp。突变体MuR6-TR与pET32a连接，并通过酶切和测序进行验证。将重组蛋白MuR6-TR转化并在大肠杆菌中表达。CCK-8实验结果显示，MuR6-TR抑制BxPC-3和PANC-1细胞的生长呈剂量依赖性，其IC50值分别为4.63和7.84 ng/ml，远低于天然TRAIL。MuR6-TR对肿瘤生长的抑制作用（24.2%）高于天然TRAIL(14.4%)，早期效果与吉西他滨相似。因此，突变体MuR6-TR在体内和体外均表现出比天然TRAIL更强的抗肿瘤作用，可能对胰腺癌具有潜在的治疗价值，有待进一步验证。
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.