吴正财, 王成艳, 吴香新, 许昌声, 林敏辉. NPM1 A型突变体对TGF-β1诱导K562细胞增殖及AKT磷酸化的影响[J]. 中国肿瘤临床, 2019, 46(4): 164-168. DOI: 10.3969/j.issn.1000-8179.2019.04.319
引用本文: 吴正财, 王成艳, 吴香新, 许昌声, 林敏辉. NPM1 A型突变体对TGF-β1诱导K562细胞增殖及AKT磷酸化的影响[J]. 中国肿瘤临床, 2019, 46(4): 164-168. DOI: 10.3969/j.issn.1000-8179.2019.04.319
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Wu Zhengcai, Wang Chengyan, Wu Xiangxin, Xu Changsheng, Lin Minhui. Effect of NPM1 mutant A on TGF-β1- induced K562 cell proliferation and AKT phosphorylation[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2019, 46(4): 164-168. DOI: 10.3969/j.issn.1000-8179.2019.04.319
Citation: Wu Zhengcai, Wang Chengyan, Wu Xiangxin, Xu Changsheng, Lin Minhui. Effect of NPM1 mutant A on TGF-β1- induced K562 cell proliferation and AKT phosphorylation[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2019, 46(4): 164-168. DOI: 10.3969/j.issn.1000-8179.2019.04.319
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NPM1 A型突变体对TGF-β1诱导K562细胞增殖及AKT磷酸化的影响

Effect of NPM1 mutant A on TGF-β1- induced K562 cell proliferation and AKT phosphorylation

    摘要
  • 摘要:
      目的  观察核仁磷酸蛋白（nucleophosmin 1，NPM1）A型突变体过表达对TGF-β1诱导的K562细胞增殖及AKT磷酸化的影响。
      方法  腺病毒载体（Ad5－NPM1）转染髓系白血病K562细胞建立过表达NPM1蛋白细胞株；应用Western blot法及ELISA法分别检测细胞NPM1蛋白表达及上清液含量，Western blot法检测K562细胞NPM1、AKT和P-AKT蛋白表达，MTT法检测细胞增殖。
      结果  K562细胞NPM1蛋白表达水平呈Ad5-NPM1转染复数（multiplicity of infection，MOI）：30-200依赖性增加，与空载组（Ad5-vector-100）相比，Ad5-NPM1-30、Ad5-NPM1-100组K562细胞及细胞上清液NPM1蛋白水平均显著增高（P < 0.01）。TGF-β1（10 ng/mL）能够诱导K562细胞AKT蛋白磷酸化，但对总AKT水平无明显影响。TGF-β1（10 ng/mL）+Ad5-NPM1-100组K562细胞P-AKT水平显著高于TGF-β1处理组（P < 0.05），各组总AKT水平无显著差别（P>0.05）。与空白对照组（CT）相比，TGF-β1（10 ng/mL）处理可促进K562细胞增殖；但TGF-β1（10 ng/mL）+Ad5-NPM1-100组细胞的增殖水平明显高于TGF-β1处理组（P < 0.01）。
      结论  NPM1可促进TGF-β1诱导的K562细胞增殖。NPM1促进TGF-β1诱导的K562细胞可能与AKT磷酸化有关。

     

    Abstract:
      Objective  To investigate the effect of nucleophosmin 1 (NPM1) mutant A on TGF-β1-induced K562 cell proliferation and AKT phosphorylation.
      Methods  K562 cells were infected with Ad5-NPM1 to create an NPM1 over-expression cell model. NPM1 levels were determined by ELISA and Western blot analysis. The levels of AKT and P-AKT were determined by Western blot. MTT was used to measure the proliferation of K562 cells.
      Results  NPM1 protein levels in K562 cells increased in an Ad5-NPM1-MOI-dependent manner. Cell proliferation and NPM1 levels in the supernatant were significantly increased in K562 cells infected with Ad5-NPM1-30 and Ad5- NPM1-100 compared to those infected with Ad5-vector-100 (P < 0.01). Treatment with (10 ng/mL) TGF-β1 increased P-AKT levels, but not total AKT levels in K562 cells. TGF-β1-induced phosphorylation of AKT was significantly increased by infection of K562 cells with Ad5-NPM1- 100. No significant differences were found in total AKT levels among all groups. TGF- β1 (10 ng/mL) treatment also increased the proliferation of K562 cells. TGF-β1-induced K562 cell proliferation was significantly increased by infection with Ad5-NPM1- 100 (P < 0.01).
      Conclusions  NPM1 improves TGF-β1-induced cell proliferation by up-regulating AKT phosphorylation levels.

     

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