Cheng Jiliang, Fang Zhaoxiang, Yang Huanhuan, Li Yong, Tian Haishan, Gong Weiyue, Chen Taotao, Liu Min, Li Xiaokun, Jiang Chao
a School of Pharmaceutical Sciences , Wenzhou Medical University , Wenzhou , China.
b College of Life and Environmental Science , Wenzhou University , Wenzhou , Zhejiang , China.
Prep Biochem Biotechnol. 2017 Aug 9;47(7):720-729. doi: 10.1080/10826068.2017.1315599. Epub 2017 Apr 14.
Fibroblast growth factor-16 (FGF16) is a member of FGF9 subfamily, which plays key role in promoting mitosis and cell survival, and also involved in embryonic development, cell growth, tissue repair, morphogenesis, tumor growth, and invasion. However, the successful high-yield purification of recombinant human fibroblast growth factor-16 (rhFGF16) protein has not been reported. In addition, lung cancer is a major cause of cancer-related deaths, which threats people's lives and its incidence has continued to rise. Learning pathways or proteins, which involved in lung tumor progression will contribute to the development of early diagnosis and targeted therapy. FGF16 promoted proliferation and invasion behavior of SKOV-3 ovarian cancer cells, whose function may be similar in lung cancer. The hFGF16 was cloned into pET-3d and expressed in Escherichia coli BL21 (DE3) pLysS. Finally, obtained two forms of FGF16 that exhibited remarkable biological activity and the purity is over 95%, meanwhile, the yield of soluble 130 mg/100 g and insoluble 240 mg/100 g. Experiments demonstrated FGF16 could promote proliferation of NCL-H460 cells by activating Akt, Erk1/2, and p38 MAPK signaling, whereas JNK had no significant effect. In total, this optimized expression strategy enables significant quantity and activity of rhFGF16, thereby meeting its further pharmacological and clinical usages.
成纤维细胞生长因子16（FGF16）是FGF9亚家族的成员，在促进有丝分裂和细胞存活中起关键作用，还参与胚胎发育、细胞生长、组织修复、形态发生、肿瘤生长和侵袭。然而，重组人成纤维细胞生长因子16（rhFGF16）蛋白的成功高产纯化尚未见报道。此外，肺癌是癌症相关死亡的主要原因，威胁着人们的生命，其发病率持续上升。了解参与肺肿瘤进展的信号通路或蛋白质将有助于早期诊断和靶向治疗的发展。FGF16促进了SKOV-3卵巢癌细胞的增殖和侵袭行为，其在肺癌中的功能可能相似。将hFGF16克隆到pET-3d中并在大肠杆菌BL21（DE3）pLysS中表达。最终，获得了两种形式的FGF16，它们表现出显著的生物学活性，纯度超过95%，同时，可溶性形式的产量为130mg/100g，不溶性形式的产量为240mg/100g。实验表明，FGF16可通过激活Akt、Erk1/2和p38 MAPK信号通路促进NCL-H460细胞的增殖，而JNK无显著影响。总之，这种优化的表达策略能够获得大量有活性的rhFGF16，从而满足其进一步的药理学和临床应用需求。
