文章摘要
刘 莹,朱祖安,崔 涛.4E-BP1?p70S6K 在RAD001 抑制胃癌MKN-28?N-87细胞生长中作用的实验研究[J].南京医科大学学报,2009,29(4):504~507
4E-BP1?p70S6K 在RAD001 抑制胃癌MKN-28?N-87细胞生长中作用的实验研究
The empirical study of the role of the expression of 4E-BP1,p70S6K in RAD001 inhibition on gastric carcinoma cell lines MKN-28, N-87
投稿时间:2008-11-10  
DOI:10.7655
中文关键词: mTOR  MKN-28  N-87  p70S6k  p-p70S6k  4E-BP1  p-4EBP1
英文关键词: mTOR  MKN-28  N-87  p70S6k  p-p70S6k  4E-BP1  p-4E-BP1
基金项目:
作者单位
刘 莹 徐州医学院病理教研室,江苏 徐州 221002 
朱祖安 徐州医学院附属医院消化内科,江苏 南京 221002 
崔 涛 徐州医学院病理教研室,江苏 徐州 221002 
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中文摘要:
      目的:探讨真核起始因子4E(eIF-4E)结合蛋白1(4E-BP1)?核糖体40S小亚基S6K蛋白激酶p70S6K 在RAD001抑制胃癌MKN-28?N-87细胞生长中的作用?方法:体外培养人胃癌MKN-28?N-87细胞,给予RAD001干预后通过细胞计数?流式细胞术?Western-blot检测肿瘤细胞生长?细胞周期分布及p70S6k?4E-BP1蛋白表达及磷酸化情况?结果:细胞计数结果显示RAD001作用于胃癌细胞株MKN-28,N-87 24 h即开始出现对细胞生长的抑制作用,随着作用时间延长至48?72 h,抑制作用逐渐增强?流式细胞术结果显示RAD001作用24 h后MKN-28?N-87细胞株细胞周期发生改变,停滞在G0~G1期细胞比例增加,在实验中显示细胞系N87对于RAD001作用相对敏感?Western-blot结果显示RAD001作用24 h后MKN-28?N-87的4E-BP1和p70S6K表达下降,同时去磷酸化?结论:RAD001是通过抑制mTOR 的活性,进而阻断4E-BP1 及p70S6K的磷酸化和蛋白质翻译进而调节着细胞周期以发挥抑制胃癌细胞生长的作用?
英文摘要:
      Objective:To investigate the role of the expression of 4E-BP1,p70S6K in RAD001 inhibition on gastric carcinoma cell lines MKN-28, N-87 and to investigate the possible mechanism. Methods:Gastric carcinoma cell lines were cultured in RPMI1640 with 10% fetal calf serum and treated by RAD001(20 nmol/L), then the cell coulter, flow cytometry, western-blot array were used to detect the growth, cell cycle and 4E-BP1,p70S6K protein expressions of the tumor cells. Results:After being treated by RAD001, the gastric carcinoma cells were inhibited in a time-dependent manner. And the percentage of Go phase was increased in MKN-28,N-87 cell lines. Meanwhile, the expressions of 4E-BP1, p70S6K, p-4E-BP1, p-p70S6K are decreased. Conclusion:The results suggested that RAD001 can inhibit the growth of gastric carcinoma cell by the adjustment of cell cycle via inhibition of mTOR, and phosphorylation of p70 S6K and 4E-BP1.
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