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背景与目的:Sep15是1998年发现的一种新的硒蛋白。文献报道Sep15可能与肿瘤的发生有关,并参与二硫键的还原。但迄今为止,Sep15的确切功能还不清楚。本实验拟对Sep15与肿瘤的关系及其抗氧化功能进行初步研究。方法:应用RT-PCR方法获得人Sep15的全长cDNA基因,并重组到真核表达载体pcDNA3.1(+)上,将重组质粒pcDNA3.1-Sep15转染到人肝癌细胞株BEL-7402,得到Sep15高表达的细胞株BEL-7402-Sep15,然后通过一系列体内和体外实验研究Sep15与肝癌细胞的关系及其抗氧化作用。结果:转染Sep15基因不影响肝癌细胞株BEL-7402的细胞形态、生长速率、克隆形成能力和裸鼠体内肿瘤生长速率;检测不同浓度外源性H2O2氧应激处理后3种细胞BEL-7402-Sep15、BEL-7402-pcDNA3.1和BEL-7402的细胞存活率,结果显示BEL-7402-Sep15的细胞存活率明显高于BEL-7402-pcDNA3.1和BEL-7402(P<0.05)。结论:转染Sep15基因的肝癌细胞株BEL-7402具有抗氧化的生物学活性。
Background and purpose: Sep15 is a new selenoprotein discovered in 1998. It has been reported in the literature that Sep15 may be involved in tumorigenesis and is involved in the reduction of disulfide bonds. But so far, the exact function of Sep15 is unclear. In this experiment, the relationship between Sep15 and tumor and their anti-oxidative function were studied. Methods: The full length cDNA of human Sep15 was obtained by RT-PCR and cloned into eukaryotic expression vector pcDNA3.1 (+). The recombinant plasmid pcDNA3.1-Sep15 was transfected into human hepatocellular carcinoma cell line BEL-7402. Sep15 high expression cell line BEL-7402-Sep15 was obtained, and then a series of in vivo and in vitro experiments were carried out to investigate the relationship between Sep15 and hepatoma cells and its anti-oxidative effect. Results: Transfection of Sep15 gene did not affect cell morphology, growth rate, clonogenic capacity and tumor growth rate in vivo in BEL-7402 hepatocellular carcinoma cell line. The expression of BEL-7402 The survival rates of BEL-7402-Sep15 cells were significantly higher than that of BEL-7402-pcDNA3.1 and BEL-7402 cells (P <0.05). Conclusion: The Bcl-7402 hepatoma cell line transfected with Sep15 gene has the biological activity of anti-oxidation.