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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 oral gland of Lampreria japonica, which is rich in cysteine and has an RGD (Arg-Gly-Asp) motif. Previous work showed that rLj-RGD1 could inhibit platelet aggregation And RGD toxin proteins typical of angiogenesis.In order to investigate whether rLj-RGD1 has another typical anti-tumor function of RGD toxin protein, the activity of rLj-RGD1 was studied in HepG2 cells of human hepatocellular carcinoma.MTT assay Showed that rLj-RGD1 inhibited the proliferation of HepG2 cells in a dose-dependent manner with an IC50 of 3.6μmol / L. The results of cell migration and invasion showed that rLj-RGD1 could inhibit HepG2 cells in a dose-dependent manner Basic fibroblast growth factor (b FGF) induced migration and infiltration.Hoechst staining and DNA Ladder apoptosis experiments showed that r Lj-RGD1 can induce apoptosis of Hep G2 cells in a dose-dependent manner.Cell adhesion experiments showed that, r Lj-RGD1 inhibits the adhesion of Hep G2 cells to vitronectin (VN) in a dose-dependent manner.The above results show that r Lj-RGD1 can inhibit the proliferation, migration and invasion of Hep G2 cells and induce it The occurrence of anoikis The results suggest that rLj-RGD1 has the typical anti-tumor function of RGD toxin protein and has the potential of becoming an antitumor drug in the future.