Identification of the sAPRIL Binding Peptide and Its Growth Inhibition Effects in the Colorectal Cancer Cells
Identification of the sAPRIL Binding Peptide and Its Growth Inhibition Effects in the Colorectal Cancer Cells
复制标题
DOI：
10.1371/journal.pone.0120564
复制
发表时间：
2015-03-31
期刊：
影响因子：
3.7
通讯作者：
He, Mei-rong
中科院分区：
文献类型：
--
作者：
He, Xiao-qing;Guan, Jing;He, Mei-rong
增殖诱导配体（APRIL）是肿瘤坏死因子（TNF）超家族成员之一。它与其特异性受体结合，参与肿瘤发生和肿瘤细胞增殖的多个过程。APRIL高表达与大肠癌的生长、转移及5-FU耐药密切相关。本研究的目的是确定一个特定的APRIL结合肽（BP）能够阻断APRIL的活性，可以作为一个潜在的治疗结直肠cancer.MethodsA噬菌体展示库被用来识别肽，选择性结合可溶性重组人APRIL（sAPRIL）。使用ELISA鉴定对sAPRIL具有最高结合亲和力的肽。分别采用CCK-8法和流式细胞术检测sAPRIL-BP对体外培养的细胞增殖和细胞周期/凋亡的影响。一个在体内的小鼠模型的结直肠癌被用来确定的sAPRIL-BP的抗肿瘤疗效。ResultsAPRIL-BP。选择具有最高亲和力的肽用于进一步表征。所鉴定的sAPRIL-BP以剂量依赖性方式抑制LOVO细胞中的肿瘤细胞增殖和细胞周期进展。在体内小鼠结肠直肠攻击模型中，sAPRIL-BP通过抑制肿瘤内增殖和诱导细胞凋亡来减少裸鼠中肿瘤异种移植物的生长。此外，在体内转移模型中，sAPRIL-BP减少结直肠癌cells.ConclusionssAPRIL-BP显着抑制肿瘤生长在体外和体内，可能是一个候选人用于治疗结直肠癌表达高水平的APRIL。
BackgroundA proliferation-inducing ligand (APRIL) is a member of the tumor necrosis factor (TNF) super family. It binds to its specific receptors and is involved in multiple processes during tumorigenesis and tumor cells proliferation. High levels of APRIL expression are closely correlated to the growth, metastasis, and 5-FU drug resistance of colorectal cancer. The aim of this study was to identify a specific APRIL binding peptide (BP) able to block APRIL activity that could be used as a potential treatment for colorectal cancer.MethodsA phage display library was used to identify peptides that bound selectively to soluble recombinant human APRIL (sAPRIL). The peptides with the highest binding affinity for sAPRIL were identified using ELISA. The effects of sAPRIL-BP on cell proliferation and cell cycle/apoptosis in vitro were evaluated using the CCK-8 assay and flow cytometry, respectively. An in vivo mouse model of colorectal cancer was used to determine the anti-tumor efficacy of the sAPRIL-BP.ResultsThree candidate peptides were characterized from eight phage clones with high binding affinity for sAPRIL. The peptide with the highest affinity was selected for further characterization. The identified sAPRIL-BP suppressed tumor cell proliferation and cell cycle progression in LOVO cells in a dose-dependent manner. In vivo in a mouse colorectal challenge model, the sAPRIL-BP reduced the growth of tumor xenografts in nude mice by inhibiting proliferation and inducing apoptosis intratumorally. Moreover, in an in vivo metastasis model, sAPRIL-BP reduced liver metastasis of colorectal cancer cells.ConclusionssAPRIL-BP significantly suppressed tumor growth in vitro and in vivo and might be a candidate for treating colorectal cancers that express high levels of APRIL.