论文部分内容阅读
为研究突变体r Lj-112蛋白的抗肿瘤活性,人工合成七鳃鳗野生型r Lj-RGD3蛋白和突变型r Lj-112蛋白,通过比较两种蛋白质的抗增殖、迁移和促凋亡的活性,确定突变体r Lj-112蛋白的生物学意义及地位。采用MTT方法检测不同浓度的r Lj-112蛋白对He La细胞增殖的抑制作用。结果表明,r Lj-112蛋白能显著抑制He La细胞的增殖,且IC50为4.3μmol/L。使用Transwell细胞培养板对b FGF诱导的He La细胞迁移实验表明,r Lj-112蛋白能抑制He La细胞的迁移。r Lj-112蛋白作用后,He La细胞经Hoechst33258和Annexin V-FITC染色结果显示,细胞均凋亡。流式细胞仪进一步证明,r Lj-112蛋白能诱导He La细胞发生凋亡,且呈剂量依赖性。由此可见,与野生型r Lj-RGD3蛋白比较,突变型r Lj-112蛋白有较高的细胞毒性作用,具有抗肿瘤的功能,有望应用于抗肿瘤基因工程药物的开发,具有重要的生物学意义。
In order to study the anti-tumor activity of the mutant rLj-112 protein, wild-type rLj-RGD3 protein and mutant rLj-112 protein were synthesized by comparing the antiproliferation, migration and apoptosis of two proteins Activity, to determine the biological significance and status of mutant r Lj-112 protein. The inhibitory effect of different concentrations of r Lj-112 protein on the proliferation of He La cells was detected by MTT assay. The results showed that rLj-112 protein could significantly inhibit the proliferation of HeLa cells with IC50 of 4.3μmol / L. Transfection of bFGF-induced HeLa cells using Transwell cell culture plates showed that rLj-112 protein inhibited HeLa cell migration. The results of Hoechst33258 staining and Annexin V-FITC staining showed that all the cells were apoptotic after rLj-112 protein treatment. Flow cytometry further demonstrated that rLj-112 protein induced HeLa cell apoptosis in a dose-dependent manner. Thus, compared with wild-type rLj-RGD3 protein, mutant rLj-112 protein has a higher cytotoxicity, with anti-tumor function, is expected to be used in the development of anti-tumor genetically engineered drugs, with important biological Significance of learning.