重组七鳃鳗富含半胱氨酸RGD蛋白rLj-RGD1对人肝癌HepG2细胞的抑制作用  

Cysteine-rich Protein rLj-RGD1 from Lampetra japonica Inhibited the Growth and Mobility of HepG2 Human Hepatocellular Carcinoma Cells

作　　者：张安伟[1] 张丕桥[1,2] 吕莉[1,3] 勾萌[1] 王继红[1] 李庆伟[1] 

机构地区：[1]辽宁师范大学生命科学学院,辽宁大连116029 [2]辽宁出入境检验检疫局,辽宁大连116001 [3]大连医科大学药学院药理教研室,大连116044

出　　处：《中国生物化学与分子生物学报》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)~~

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.

关 键 词：日本七鳃鳗 RGD毒素蛋白 抗肿瘤 迁移与浸润 失巢凋亡 

分 类 号：Q781[生物学—分子生物学]