Su Shuonan, Xu Huanli, Chen Xiaoliang, Qiao Gan, Farooqi Ammad A, Tian Ye, Yuan Ru, Liu Xiaohui, Li Cong, Li Xiao, Wu Ning, Lin Xiukun
Department of Pharmacology, Capital Medical University, Beijing 100069, China.
School of medicine, Shanxi Datong University, Datong 037009, China.
Recent Pat Anticancer Drug Discov. 2018;13(3):360-367. doi: 10.2174/1574892813666180628170240.
CS5931, a novel marine peptide, was extracted and purified from the sea squirt Ciona savignyi. Our previous studies showed that recombinant CS5931 can significantly inhibit tumor growth both in vitro and in vivo. However, its molecular targets have not been elucidated.
The target of the recombinant CS5931 was identified by pull-down/SDS-PAGE/MS approaches and confirmed by Western blot and surface plasmon resonance analysis. Transwell experiments were used to detect whether the recombinant CS5931 inhibited cancer migration and invasion via enolase 1. Dot blotting analysis was used to detect the effect of CS5931 on the interaction of enolase 1 and plasminogen, as well as enolase 1 and uPA/uPAR.
Enolase 1 was identified as the molecular target interacting with the recombinant CS5931. Transwell experiment showed that the recombinant CS5931 was able to inhibit migration and invasion of HCT116 cells and enolase 1 overexpression reversed the effects of the recombinant CS5931 on migration and invasion of cancer cells. Dot blotting analysis revealed that the recombinant CS5931 interfered with the interaction among enolase 1 and plasminogen as well as enolase 1 and uPA/uPAR.
Our present study showed that the recombinant CS5931 could inhibit tumor invasion and matastasis via interacting with enolase 1, suggesting that the new marine polypeptide CS5931 possesses the potential to be developed as a novel anticancer agent.
CS5931是一种从海鞘萨氏海鞘中提取纯化的新型海洋肽。我们之前的研究表明,重组CS5931在体外和体内均能显著抑制肿瘤生长。然而,其分子靶点尚未阐明。
通过下拉/SDS-PAGE/MS方法鉴定重组CS5931的靶点,并通过蛋白质印迹和表面等离子体共振分析进行确认。采用Transwell实验检测重组CS5931是否通过烯醇化酶1抑制癌症迁移和侵袭。斑点印迹分析用于检测CS5931对烯醇化酶1与纤溶酶原以及烯醇化酶1与尿激酶型纤溶酶原激活剂/尿激酶型纤溶酶原激活剂受体相互作用的影响。
烯醇化酶1被鉴定为与重组CS5931相互作用的分子靶点。Transwell实验表明,重组CS5931能够抑制HCT116细胞的迁移和侵袭,烯醇化酶1过表达可逆转重组CS5931对癌细胞迁移和侵袭的影响。斑点印迹分析显示,重组CS5931干扰了烯醇化酶1与纤溶酶原以及烯醇化酶1与尿激酶型纤溶酶原激活剂/尿激酶型纤溶酶原激活剂受体之间的相互作用。
我们目前的研究表明,重组CS5931可通过与烯醇化酶1相互作用抑制肿瘤侵袭和转移,提示新型海洋多肽CS5931具有开发成为新型抗癌药物的潜力。