摘要
目的探讨SGK3基因对乳腺癌细胞侵袭转移能力的影响。方法构建pEGFP-N1-SGK3重组质粒,用载体质粒pEGFP-N1和重组质粒pEGFP-N1-SGK3转染乳腺癌MCF7细胞;MTT法观察转染细胞的增殖情况,Tran-swell细胞迁移实验检测SGK3过表达对细胞迁移能力影响,RT-PCR检测相关基因表达,Western blot检测SGK3过表达对Wnt/β-catenin信号通路关键因子的影响。结果利用pEGFP-N1-SGK3质粒转染乳腺癌MCF7细胞,建立表达SGK3蛋白的细胞系;细胞增殖实验发现,SGK3的过表达使MCF7细胞生长速度加快;细胞迁移实验发现过表达SGK3的细胞运动迁移能力增强,其可使mmp9的表达增强,而brms1表达无明显变化;Western blot结果显示SGK3过表达可增加乳腺癌细胞磷酸化的GSK3β水平。结论 SGK3的过表达通过影响Wnt/β-catenin信号通路中的GSK3β磷酸化而增强细胞迁移能力。
Objective To investigate the effects of overexpression of serum and glucocorticoid-regulated protein kinase 3（SGK3） on invasive and metastatic ability of breast cancer cell.Methods Breast cancer cell（MCF7） were transfected respectively by carrier plamid（pEGFP-N1） and recombinant plasmid（pEGFP-N1-SGK3） after recombinant plasmid（pEGFP-N1-SGK3） were constructed;The cell multiplication were detected by MTT,Effects of overexpression of SGK3 on cell migration were examined using transwell cell migration experiment;RT-PCR were used to measured related gene;effects of overexpression of SGK3 on key factor of Wnt/β-catenin signal path were detected by Western blot.Results The cell line of SGK3 protein were constructed after transfection with recombinant plasmid（pEGFP-N1-SGK3）.Our study showed that the growth velocity of MCF7 was significantly faster than before after overexpression of SGK3.As compared with normal MCF7 cell,the ability of cell migration was more stronger,and the expression of mmp9 was strengthened for this reason,but no change of expression of brmsl was observed.The level of GSK3β phosphorylation was enhanced by overexpression of SGK3.Conclusion The results showed that the overexpression of SGK3 could strengthen cell migration through effected on GSK3β phosphorylation in Wnt/β-catenin signal path.
出处
《中国实验诊断学》
2012年第3期391-395,共5页
Chinese Journal of Laboratory Diagnosis
基金
黑龙江省教育厅科研项目(11531431)
齐齐哈尔市科技局指导项目
