过表达Sprouty2分子促进HEK293T细胞增殖与存活

Over-expression of human SPRY2 promotes the proliferation and survival of HEK293T cells

目的构建表达人Sprouty 2(SPRY2)基因的重组表达载体pDC315/hSPRY2,转染人胚肾(HEK)293T细胞表达目的蛋白,初步探讨其对HEK293T细胞增殖与存活的影响。方法构建hSPRY2重组腺病毒表达载体,体外转染HEK293T细胞,以Western blot法检测目的蛋白的表达,以CCK-8法检测SPRY2对HEK293T细胞增殖或去血清条件下细胞存活的影响。结果成功构建重组表达载体pDC315/hSPRY2,在转染的HEK293T细胞中检测到目的蛋白hSPRY2的表达,转染pDC315/hSPRY2组HEK293T细胞的增殖率及存活率均明显高于对照组即转染pDC315/EGFP组(P<0.05)。结论成功构建了hSPRY2重组表达载体,过表达hSPRY2可促进HEK293T细胞的增殖与存活。

Objective To construct a recombinant expression vector pDC315/hSPRY2 carrying human SPRY2 （hSPRY2） gene, and transfect the recombinant plasmid into human embryonic kidney HEK293T cells to express the target protein, and thus explore preliminarily the effect of hSPRY2 on the proliferation and survival of HEK293T cells. Methods The recombinant adenoviral vector expressing hSPRY2 was constructed and transfected into HEK293T cells in v/tro. The expression of hSPRY2 target protein in the transfected cells was identified by Western blot analysis. The effect of hSPRY2 on the proliferation and survival of HEK293T cells in a serum-free condition was tested using CCK-8 assay. Results The recombinant expression vector pDC315/hSPRY2 was constructed, and the target protein hSPRY2 was expressed in the transfected HEK293T cells. The proliferation and survival rates of HEK293T cells transfected with pDC315/hSPRY2 were significantly higher than those transfected with pDC315/EGFP （P 〈 0.05）. Conclusion The recombinant expression vector for hSPRY2 was successfully constructed and its expression was demonstrated in the transfected HEK293T cells. Over-expression of SPRY2 promoted the proliferation and survival of HEK293T cells.