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[KLF4过表达对K562细胞体外增殖和凋亡的影响]

[Effect of Over-Expression of KLF4 on the Proliferation and Apoptosis of K562 Cells In Vitro].

作者信息

Li Bo, Hu Hai-Yan, Tan Ming-Xian

机构信息

Clinical Laboratory, Three Gorges Central Hospital of Chongqing, Wanzhou 404000, Chongqing, China.

Clinical Laboratory, Three Gorges Central Hospital of Chongqing, Wanzhou 404000, Chongqing, China. E-mail:

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2017 Oct;25(5):1373-1377. doi: 10.7534/j.issn.1009-2137.2017.05.016.

DOI:10.7534/j.issn.1009-2137.2017.05.016
PMID:29070110
Abstract

OBJECTIVE

To study the effect of over-expression of KLF4 on the proliferation and apoptosis of K562 cells.

METHODS

The recombinant plasmid with over-expression of KLF4 was transfected into K562 cells by electroporation, then the obtained G418-resistant clones of K562 cells (K562/pEGFP-KLF4) were used as experimental group, and the vector control (K562/pEGFP-C1) and blank K562 cell control groups were set up at the same time. The expression of EGFP-KLF4 fusion protein in K562 cells was observed by fluorescence microscopy; the Western blot was performed to detect the level of KLF4 protein in the cells of each groups, the cell proliferation was tested by methylthiazolyl tetrazolium (MTT) assay, and the cell apoptosis was detected by flow cytometry.

RESULTS

Compared with K562/pEGFP-C1 group and blank K562 group, the level of KLF4 protein of K562/ pEGFP-KLF4 cells were significantly increased, and the over-expression ratio reached 74.07%(P<0.05). The proliferation ability of the over-expressing KLF4 K562 cells was inhibited significantly (P<0.05). However, the apoptosis of K562/pEGFP-ILK cells with over-expression of KLF4 was increased (P<0.05).

CONCLUSION

A stable over-expressing KLF4 protein K562 cell line with was constructed successfully, and the over-expression of KLF4 can inhibi proliferation and promote apoptosis of K562 cells.

摘要

目的

研究KLF4过表达对K562细胞增殖和凋亡的影响。

方法

通过电穿孔法将KLF4过表达的重组质粒转染至K562细胞,将获得的K562细胞G418抗性克隆(K562/pEGFP-KLF4)作为实验组,同时设立载体对照组(K562/pEGFP-C1)和K562细胞空白对照组。用荧光显微镜观察K562细胞中EGFP-KLF4融合蛋白的表达;采用蛋白质免疫印迹法检测各组细胞中KLF4蛋白水平,用噻唑蓝(MTT)法检测细胞增殖情况,用流式细胞术检测细胞凋亡情况。

结果

与K562/pEGFP-C1组和K562细胞空白组相比,K562/pEGFP-KLF4细胞的KLF4蛋白水平显著升高,过表达率达74.07%(P<0.05)。KLF4过表达的K562细胞增殖能力明显受到抑制(P<0.05)。然而,KLF4过表达的K562/pEGFP-ILK细胞凋亡增加(P<0.05)。

结论

成功构建了稳定过表达KLF4蛋白的K562细胞系,KLF4过表达可抑制K562细胞增殖并促进其凋亡。

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