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PDCD4对DAP5表达及大肠癌细胞凋亡的影响

易波1,李其云1,饶华民1,邵江华2   

  1. 1. 江西省肿瘤医院
    2. 南昌大学第二附属医院肝胆外科
  • 收稿日期:2012-07-11 修回日期:2013-01-10 出版日期:2013-06-15 发布日期:2013-06-15
  • 通讯作者: 易波

Effects of Programmed Cell Death 4 Gene on Expression of Death Associated Protein 5 and Apoptosis in Colon Cancer Cells

YI Bo 1,LI Qiyun 1,RAO Huamin 2,SHAO Jianghua 1   

  1. 1. Department of Abdominal Surgery，Jiangxi Province Tumor Hospital
    2. Department of Abdominal Surgery.Jiangxi Province Tumor Hospital
  • Received:2012-07-11 Revised:2013-01-10 Published:2013-06-15 Online:2013-06-15
  • Contact: YI Bo

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摘要： 【摘要】目的  探讨程序性死亡基因4(PDCD4)对死亡相关蛋白5（DAP5) 的表达及对人大肠癌细胞系LOVO凋亡的影响。方法  构建重组真核表达载体pFLAG/PDCD4，转染人大肠癌细胞LOVO，G418（500 mg/L）筛选获得稳定表达PDCD4的细胞系。RT-PCR及Western blotting检测LOVO细胞中PDCD4的mRNA及蛋白表达，流式细胞仪检测LOVO细胞凋亡情况，Western blotting检测LOVO细胞DAP5表达的变化。结果 成功建立稳定表达PDCD4的大肠癌  细胞LOVO-pFLAG/PDCD4。转染PDCD4 的LOVO-pFLAG/PDCD4 组与空白对照LOVO 组、转染空载质粒的LOVO-pFLAG组相比，PDCD4蛋白表达和mRNA水平明显升高（P < 0.01）；细胞凋亡率明显增加（P < 0.01）；同时伴有DAP5蛋白表达明显升高（P < 0.01）。结论PDCD4能够诱导大肠癌细胞LOVO的凋亡，其机制可能与上调DAP5的表达有关。

关键词: 蛋白激酶类, 结直肠肿瘤, 癌, 细胞凋亡, 质粒, 转染, 重组, 遗传

Abstract: [Abstract]   Objective   To investigate the effect of exogenous programmed cell death4(PDCD4) gene on the expres-
sion of death associated protein5(DAP5) and apoptosis of human colorectal cancer cell LOVO, and involved mechanisms thereof.   Methods   Recombinant eukaryotic plasmid pFLAG/PDCD4was constructed and transfected into human colorectal cancer cell LOVO. Cells stably expressing PDCD4were established by G418selection (500mg/L). The levels of PDCD4protein and mRNA were analyzed by RT-PCR and Western blotting. The apoptotic cells were measured by flow cytometry, and protein expression of DAP5was detected by Western blotting.  Results LOVO-pFLAG/PDCD4cell line was successfully es-tablished by G418selection. Compared to non-transfection and mock-transfection group, the levels of PDCD4mRNA and protein were significantly increased, the cell apoptosis ratio was enhanced and the expression of DAP5protein was increased in transfection group (P<0.01).   Conclusion PDCD4could induce apoptosis of human colorectal cancer LOVO cells, which mechanism might be involved in up-regulating DAP5protein.

Key words: protein kinases, colorectal neoplasms, carcinoma, cell cycles, Plasmids, transfention, recombination, 遗传