摘要
rLj-RGD1为源于日本七鳃鳗口腔腺的基因重组蛋白,其富含半胱氨酸并具有一个RGD(Arg-Gly-Asp)模体.前期工作表明,r Lj-RGD1具有抑制血小板聚集及血管新生的RGD毒素蛋白典型功能.为了研究rLj-RGD1是否具有RGD毒素蛋白的另一典型抗肿瘤功能,以人肝癌Hep G2细胞为模型,对r Lj-RGD1进行了活性研究.MTT法结果显示,r Lj-RGD1呈剂量依赖方式抑制HepG2细胞的增殖,其半抑制浓度(IC50)为3.6μmol/L;细胞迁移与浸润实验结果显示,r Lj-RGD1能以剂量依赖方式抑制Hep G2细胞碱性成纤维生长因子(b FGF)诱导的迁移与浸润.Hoechst染色和DNA Ladder等细胞凋亡实验表明,r Lj-RGD1能够以剂量依赖方式诱导Hep G2细胞发生凋亡.细胞黏附实验表明,r Lj-RGD1以剂量依赖方式抑制Hep G2细胞与玻连蛋白(vitronectin,VN)的黏附.上述结果表明,r Lj-RGD1具有抑制人肝癌Hep G2细胞增殖、迁移和浸润的功能,并可诱导其发生失巢凋亡.本研究结果提示,r Lj-RGD1具有典型的RGD毒素蛋白抗肿瘤功能,其未来具有成为抗肿瘤药物的潜力.
rLj-RGD1 is a recombinant protein derived from the buccal gland of Lampetra japonica,which consists of 14 cysteines and a RGD（ Arg-Gly-Asp） motif. Previous study showed that rLj-RGD1 inhibited platelet aggregation and anti-angiogenesis like other RGD toxin proteins. To explore whether r Lj-RGD1 has anti-tumor activities,HepG2 human hepatocellular carcinoma cells were used. The MTT assayrevealed that,rLj-RGD1 inhibited Hep G2 proliferation in a dose-dependent manner with IC50 at 3. 6μmol /L,as well as b FGF-induced cell migration and invasion. Hoechst staining and DNA ladder assays demonstrated that rLj-RGD1 treatments induced apoptosis of HepG2 cells with increasing doses. In addition,rLj-RGD1 was also found to significantly inhibit cell adhesion to vitronectin that could lead to anoikis. Our results demonstrated that r Lj-RGD1 possessed anti-tumor functions of typical RGD toxin proteins.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2015年第6期630-635,共6页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家高技术研究发展计划(863计划
No.SS2014AA091602)
国家海洋公益项目(No.201305016-5)
国家自然科学基金(No.30770297)
大连市重大科技攻关项目(No.2013E11SF056
No.2014E12SF067)~~
