PEP06 polypeptide 30 exerts antitumour effect in colorectal carcinoma via inhibiting epithelial-mesenchymal transition
PEP06 polypeptide 30 exerts antitumour effect in colorectal carcinoma via inhibiting epithelial-mesenchymal transition
复制标题
PEP06多肽30通过抑制上皮间质转化在结直肠癌中发挥抗肿瘤作用
DOI：
10.1111/bph.14352
复制
发表时间：
2018-08-01
影响因子：
7.3
通讯作者：
Yang, Baofeng
中科院分区：
文献类型：
--
作者：
Yu, Siming;Li, Linna;Yang, Baofeng
背景与目的研究了由内皮抑素修饰的多肽pep06对结直肠癌（CRC）的抗肿瘤作用，并在体外和体内模型中探讨了其抗肿瘤活性的可能机制。实验方法：用PEP06处理结直肠癌细胞，进行细胞增殖和迁移实验。采用Western blot、免疫荧光染色和免疫组织化学检测体外和残余异种移植物模型的上皮-间质转化（EMT）进展。通过miRNA芯片鉴定PEP06调控的miRNA，并进行原位杂交和实时荧光定量PCR验证。利用Biacore SA生物芯片检测PEP06与整合素α v β 3的相互作用。通过功能获得和功能丧失方法验证miR-146b-5p的细胞功能。采用小鼠肺转移模型研究PEP06对肺转移生长的影响。关键结果tspep06不影响细胞活力，但降低了SW620和HCT116细胞的迁移和EMT。PEP06通过与整合素α v β 3结合，显著抑制miR-146b-5p在这两种细胞系中的表达。MiR-146b-5p被证明通过靶向Smad4增加EMT， MiR-146b-5p -Smad4级联调节CRC中的EMT。PEP06还通过下调miR-146b-5p抑制EMT，抑制结直肠癌肺转移，提高小鼠存活率，阻碍残余肿瘤生长。结论与意义spep06是一种多肽，通过其RGD基序结合整合素α v β 3抑制结肠癌的生长和转移，从而下调miR-146b-5p，从而在体外和体内抑制EMT。它可能有治疗结直肠癌的潜力。
BACKGROUND AND PURPOSEPEP06, a polypeptide modified from endostatin, was investigated for its antitumour effects on colorectal cancer (CRC) and the possible mechanisms of this antitumour activity were examined in in vitro and in vivo models.EXPERIMENTAL APPROACHAfter PEP06 treatment, cell proliferation and migration assays were performed in CRC cells. Epithelial-mesenchymal transition (EMT) progression was determined by Western blotting, immunofluorescent staining and immunohistochemistry in vitro and in a residual xenograft model. MiRNAs regulated by PEP06 were identified by miRNA microarray and verified by in situ hybridization and quantitative real-time PCR. The interactions between PEP06 and integrin alpha v beta 3 were determined with Biacore SA biochips. The cellular function of miR-146b-5p was validated by gain-of-function and loss-of-function approaches. A mouse model of lung metastasis was used to determine the effect of PEP06 on metastatic growth.KEY RESULTSPEP06 did not affect cell viability but reduced migration and EMT in SW620 and HCT116 cells. PEP06 significantly repressed the expression of miR-146b-5p in these two cell lines through binding to integrin alpha v beta 3. MiR-146b-5p was shown to increase EMT by targeting Smad4, and the miR-146b-5p-Smad4 cascade regulated EMT in CRC. PEP06 also suppressed CRC pulmonary metastasis, increased survival of mice and hampered residual tumour growth by inhibiting EMT through down-regulating miR-146b-5p.CONCLUSIONS AND IMPLICATIONSPEP06 is a polypeptide that inhibits the growth and metastasis of colon cancer through its RGD motif binding to integrin alpha v beta 3, thereby down-regulating miR-146b-5p to inhibit EMT in vitro and in vivo. It might have potential as a therapeutic for CRC.