上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

腺花素通过peroxiredoxinⅠ对端粒酶表达和细胞死亡的影响

崔佳毅，曹 阳，雷 虎，徐含章，吴英理   

  1. 上海交通大学 医学院病理生理学教研室 细胞分化与凋亡教育部重点实验室， 上海 200025
  • 出版日期:2015-01-28 发布日期:2015-01-29
  • 通讯作者: 吴英理, 电子信箱: wuyingli@shsmu.edu.cn。
  • 作者简介:崔佳毅(1985—)， 女， 硕士生; 电子信箱: keyhsy@shsmu.edu.cn。
  • 基金资助:

    上海市科委资助项目(11JC1406500)；上海市教委资助项目（13YZ028）

Effects of adenanthin to cell death and expression of telomerase through peroxiredoxin Ⅰ

CUI Jia-yi, CAO Yang, LEI Hu, XU Han-zhang, WU Ying-li   

  1. The Key Laboratory of Cell Differentiation and Apoptosis of the Chinese Ministry of Education, Department of Pathophysiology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
  • Online:2015-01-28 Published:2015-01-29
  • Supported by:

    Foundation of Science and Technology Commission of Shanghai Municipality, 11JC1406500; Foundation of Shanghai Municipal Education Commission, 13YZ028

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摘要：

目的 探讨腺花素对白血病细胞端粒酶的影响和作用机制。方法 用不同浓度腺花素（0、2、4、6 μmol/L）处理NB4-LR1细胞不同时间（0、6、12、24 h），Western blotting检测人端粒酶逆转录酶(hTERT)蛋白的表达，Real-time PCR 检测hTERT mRNA水平。用RNA干扰方法在NB4-LR1细胞内降低peroxiredoxin Ⅰ （Prdx Ⅰ）的水平，Western blotting检测hTERT蛋白的表达水平。在NB4-LR1细胞内过表达hTERT，锥虫蓝拒染法检测不同浓度（2、4 μmol/L）的腺花素处理NB4-LR1-hTERT-GFP、NB4-LR1-GFP细胞后，细胞的增殖和活力的改变。结果 Western blotting检测结果显示，腺花素能够呈时间－剂量依赖性抑制hTERT的表达。敲除PrdxⅠ后hTERT的表达下降。在NB4-LR1细胞过表达hTERT可以部分抑制腺花素对细胞的毒性作用但不能抑制其诱导的细胞分化。结论 腺花素通过PrdxⅠ抑制端粒酶表达，促进细胞的死亡。

关键词: 端粒酶, peroxiredoxinⅠ, 腺花素, 白血病, 细胞死亡

Abstract:

Objective To investigate the effects of adenanthin on telomerase of leukemia NB4-LR1 cells and relevant mechanisms. Methods NB4-LR1 cells were treated with different concentrations of adenanthin (0, 2, 4, and 6 μmol/L) for different periods of time (0, 6, 12, and 24 h). The expression of human telomerase reverse transcriptase (hTERT) protein was detected by Western blotting, and the expression of hTERT mRNA was detected by Real-time PCR. The RNA interference method was adopted to decrease the expression of peroxiredoxin Ⅰ (Prdx Ⅰ) in NB4-LR1 cells. Then the expression of hTERT protein was detected by Western blotting. hTERT was over-expressed in NB4-LR1 cells and the NB4-LR1-hTERT-GFP and NB4-LR1-GFP cells were treated with different concentrations of adenanthin (2 and 4 μmol/L). The variations of proliferation and viability of cells were detected by trypan blue exclusion assay. Results The results of Western blotting showed that adenanthin inhibited the expression of hTERT in a dosetime dependent manner. The expression of hTERT decreased after Prdx I was knocked down. Over-expression of hTERT in NB4-LR1 cells partially inhibited the toxic effect of adenanthin on cells, but could not inhibit cell differentiation induced by adenanthin. Conclusion Adenanthin inhibits the expression of telomerase through Prdx Ⅰ and contributes to the death of cells.

Key words: telomerase, peroxiredoxin Ⅰ, adenanthin, leukemia, cell death