[鱼藤素在体外对U937细胞的细胞周期及核孔复合体蛋白Nup98和Nup88的调控作用]

[Deguelin regulates cell cycle and nuclear pore complex protein Nup98 and Nup88 in U937 cells in vitro].

作者信息

Chen Yan, Liu Hong-Li, Cui Guo-Hui, Wu Qiu-Ling, He Jing, Chen Wei-Hua

机构信息

Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.

出版信息

Zhonghua Xue Ye Xue Za Zhi. 2007 Feb;28(2):115-8.

PMID:17650673

Abstract

OBJECTIVE

To investigate antitumor activity and molecular mechanism of deguelin to the human U937 leukaemia cells and to explore the mechanisms regulating cell cycle and nucleoporin 98 (Nup98) and nucleoporin 88 (Nup88) in vitro.

METHODS

The effects of deguelin on the growth of U937 cells were studied by MTT assay, and the cell cycle of U937 cells by a propidium iodide method. The localization of the nuclear pore complex protein Nup98 and Nup88 was checked by immunofluorescence and immunoelectron microscopy. The expressions of Nup98 and Nup88 in U937 cells were checked by flow cytometry (FCM) and Western blot respectively.

RESULTS

The proliferation of U937 cells was significantly inhibited in a time-dose dependent manner in deguelin-treated group with a 24 h IC50 value of 21.61 nmol/L and 36 h IC50 value of 17.07 nmol/L. U937 cells treated with deguelin showed reduction in the percentages of cells in G0/G1, whereas accumulation of cells in S and G2/M phase. The ratio of G1/G0 phase cells were 73.01%, 71.15%, 68.42%, 52.45%, 43.99% and 22.82%, and that of S phase cells were 17.18%, 16.30%, 18.09%, 27.56%, 31.21% and 46.85%, and that of G2/M phase cells were 9.75%, 12.31%, 13.09%, 18.99%, 24.83% and 27.79% at deguelin concentrations of 0, 5, 10, 20, 40, 80 nmol/L respectively. Nup88 and Nup98 were found on both the nuclear and cytoplasmic side of the U937 cells. The expression of Nup98 was up-regulated and Nup88 down-regulated in deguelin treated U937 cells.

CONCLUSION

Deguelin is able to inhibit the proliferation of U937 cells by regulating the cell cycle. The antitumor activity of deguelin was related to up-regulating the expression of Nup98 and down-regulating Nup88 protein.

摘要

目的

研究鱼藤素对人U937白血病细胞的抗肿瘤活性及分子机制，体外探讨其调控细胞周期以及核孔蛋白98（Nup98）和核孔蛋白88（Nup88）的机制。

方法

采用MTT法研究鱼藤素对U937细胞生长的影响，用碘化丙啶法检测U937细胞的细胞周期。通过免疫荧光和免疫电子显微镜检查核孔复合体蛋白Nup98和Nup88的定位。分别用流式细胞术（FCM）和蛋白质免疫印迹法检测U937细胞中Nup98和Nup88的表达。

结果

鱼藤素处理组U937细胞的增殖呈时间 - 剂量依赖性显著抑制，24 h的IC50值为21.61 nmol/L，36 h的IC50值为17.07 nmol/L。用鱼藤素处理的U937细胞G0/G1期细胞百分比降低，而S期和G2/M期细胞积累。在鱼藤素浓度分别为0、5、10、20、40、80 nmol/L时，G1/G0期细胞比例分别为73.01%、71.15%、68.42%、52.45%、43.99%和22.82%，S期细胞比例分别为17.18%、16.30%、18.09%、27.56%、31.21%和46.85%，G2/M期细胞比例分别为9.75%、12.31%、13.09%、18.99%、24.83%和27.79%。在U937细胞的细胞核和细胞质中均发现Nup88和Nup98。在鱼藤素处理的U937细胞中，Nup98的表达上调，Nup88的表达下调。