胃癌细胞MKN-45和SGC-7901中MTA1的泛素化修饰及其对MTA1表达的影响
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国家自然科学基金(81272734,81372588)


Ubiquitination and expression of MTA1 in gastric cancer MKN-45 and SGC-7901 cells
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    摘要:

    目的:转移相关基因1(metastasis associated 1,MTA1)已知与肿瘤转移密切相关,且与胃癌恶性程度明显相关,并可以通过影响信号转导途径发挥其功能,本研究拟确定MTA1是否可以被泛素化途径所调控-方法:以蛋白酶体抑制剂MG-132处理胃癌细胞后,以IP/WB实验检测MTA1的表达水平,并将带Myc标签的MTA1质粒以及带HA标签的泛素(HA-Ub)质粒转入该细胞,检测Myc-MTA1的表达-结果:以蛋白酶体抑制剂MG-132处理胃癌细胞MKN-45-SGC-7901后,MTA1表达水平显著增加;免疫共沉淀实验显示泛素分子可以与MTA1直接结合,并且实验组较对照组相比泛素化程度明显增加;将HEK293细胞以siMTA1处理封闭MTA1的表达后,将Myc-MTA1质粒以及HA-Ub质粒转入该细胞,发现当后者存在时Myc-MTA1可被强烈泛素化-结论:在胃癌细胞中MTA1是可被泛素化途径修饰的蛋白-

    Abstract:

    Objective:Metastasis associated 1 (MTA1)is a protein closely related to the malignant degree of gastric cancer. We try to prove that MTA1 can be regulated by the ubiquitin pathway in this study. Methods:After the proteasome inhibitor MG-132 treatment in gastric cancer cells,the expression level of MTA1 was detected though IP/WB. MTA1 expressing plasmid containing the Myc tag and HAtagged ubiquitin plasmid (HA-Ub) were co-transfected into cells,and the expression of Myc-MTA1 was detected. Results:After treatment with MG-132,the protein level of MTA1 in gastric carcinoma cell line SGC-7901 increased significantly. Myc-MTA1 was strongly ubiquitinated by HA-Ub in HEK293 cells. Conclusion:MTA1 in human gastric cancer cells can be regulated through ubiquitination pathway.

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付海京,王 锐,陈一天,张 群,孙 茜,黄小银,管晓翔,王靖华,陈龙邦.胃癌细胞MKN-45和SGC-7901中MTA1的泛素化修饰及其对MTA1表达的影响[J].南京医科大学学报(自然科学版),2014,(11):1503-1506

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  • 收稿日期:2013-04-02
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  • 在线发布日期: 2014-11-21
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