天津医药

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Akt1和Akt2基因转染对人胃黏膜上皮细胞GES-1侵袭和迁移的影响

谢甲贝1,张庆瑜2,康春生3,4,韩静2,4,王涛2,4,张洁5   

  1. 1. 天津医科大学总医院消化内科
    2. 天津医科大学总医院消化科
    3. 天津医科大学总医院神经病学研究所
    4.
    5. 天津医科大学总医院
  • 收稿日期:2010-11-23 修回日期:2010-12-14 出版日期:2011-04-15 发布日期:2011-04-15
  • 通讯作者: 张庆瑜

The effect of Akt1 and Akt2 gene transfection on invasion and migration of human gastric epithelial cell GES-1

  • Received:2010-11-23 Revised:2010-12-14 Published:2011-04-15 Online:2011-04-15

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摘要： 【摘要】目的 观察蛋白激酶B1（Akt1）和蛋白激酶B2（Akt2）基因脂质体法转染人胃黏膜上皮细胞系GES-1后对其侵袭和迁移的影响。 方法 采用脂质体法将分别含 Akt1和 Akt2基因全长 cDNA 序列的p-LXSN-Akt1（Akt1组）和p-LXSN-Akt2 （Akt2组）质粒转染人胃黏膜上皮细胞系GES-1，以转染p-LXSN空载质粒的细胞为空载组，以正常未转染细胞为对照组，并经抗生素G418筛选抗性克隆，蛋白印迹（Western blot）鉴定各组细胞Akt1,、Akt2及基质金属蛋白酶2(MMP-2)的蛋白表达水平；Transwell法分析转然后细胞的侵袭能力；用免疫荧光法观察转染后GES-1细胞丝状肌动蛋白（F-actin）的变化。结果 ①Akt1和Akt2组均获得了稳定表达Akt1和Akt2基因的GES-1细胞模型；两组中Mmp-2蛋白表达均高于空载组和对照组（P<0.01）；②Transwell法显示Akt1和Akt2组GES-1细胞侵袭能力均明显增加（P<0.01）；③免疫荧光发现Akt1和Akt2组细胞丝状肌动蛋白F-actin的均聚集增加。结论 Akt1、Akt2基因均可以通过增加细胞侵袭力和迁移力，促进GES-1细胞恶性转化

关键词: Akt1基因, Akt2基因, GES-1, 侵袭, 迁移

Abstract: The effect of Akt1 and Akt2 gene transfection on invasion and migration of human gastric epithelial cell GES-1 XIE jia-bei, ZHANG Qing-yu*, HAN jing ,WANG tao, ZHANG jie. Department of Gastroenterology, Tianjin Medical University General Hospital, Tianjin 300052,China Kang Chun-sheng, Department of Neurosurgery, Tianjin Medical University General Hospital and Laboratory of Neuro-Oncology, Tianjin Neurological Institute, Tianjin 300052,China. Corresponding author : ZHANG Qing-yu, Email: zhangqy@tijmu.edu.cn Abstract To observe the effects of Akt1 and Akt2 gene transfections on invasion and migration of human gastric epithelial cell line GES-1. Methods The plasmid of p-LXSN-Akt1 which contains the whole Akt1 gene sequence and p-LXSN-Akt2 which contains the whole Akt2 gene sequence were used to transfect GES-1 cells with lipofectamine, The GES-1 cells which were transfected by the empty vector p-LXSN were grouped as the black vector ,the no transfected ones were the control. Then the positive cell clones were selected with antibiotics G418. The protein expressions of Akt1, Akt2 and Mmp-2 were measured by Western blotting analysis; and Transwell assay analyzed their invasion and metastasis capability. The expressions of F-actin were detected by Immunofluorescence. Results Cell modes which expresses Akt1 gene (Akt1 group) and which expresses Akt2 gene (Akt2 group) were obtained; Compared with the black vector group and control group, The protein expressions of Mmp-2 in Akt1 and Akt2 groups were increased significantly.（P<0.01; transwell assay showed Akt1 and Akt2 groups could significantly advance the invasion and metastasis of cells; the expressions of F-actin were increased corresponding1y in Akt1 and Akt2 groups. Conclusion Akt1, Akt2 gene could increase cell invasion and migration, which showed the promotion of malignant transformation of GES-1 cells .

Key words: Akt1 gene, Akt2 gene, GES-1, invasion, migration