The effect of hnRNP K shRNA on K562 cells

HUANG Lian-fen; et al

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The effect of hnRNP K shRNA on K562 cells

更新时间：2024-01-17

- The effect of hnRNP K shRNA on K562 cells
- Journal of Chongqing Medical University Issue 9, Pages: 1071-1075(2008)
- 作者机构：
  
  重庆医科大学医学检验系临床检验诊断学省部共建教育部重点实验室
- 作者简介：
- 基金信息：
- DOI：
  CLC： R733.7
- Published：2008，
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[1]黄莲芬,冯文莉,罗宏伟,孙成铭.hnRNP K shRNA对K562细胞效应的初步研究[J].重庆医科大学学报,2008(09):1071-1075+1147.

HUANG Lian-fen, et al. The effect of hnRNP K shRNA on K562 cells. [J]. Journal of Chongqing Medical University (9):1071-1075(2008)
[1]黄莲芬,冯文莉,罗宏伟,孙成铭.hnRNP K shRNA对K562细胞效应的初步研究[J].重庆医科大学学报,2008(09):1071-1075+1147. DOI：

HUANG Lian-fen, et al. The effect of hnRNP K shRNA on K562 cells. [J]. Journal of Chongqing Medical University (9):1071-1075(2008) DOI：

摘要

目的:探索RNA结合蛋白K（Heterogeneous nuclear ribonucleoprotein K

hnRNP K）shRNA逆转录病毒对K562细胞的效应。方法:HindⅢ/EcorⅠ双酶切和测序鉴定pSUPER retro hnRNP K shRNA逆转录病毒载体

用包装得到的逆转录病毒感染K562细胞

G418稳定筛选后

分别用流式细胞仪

MTT实验

瑞氏染色

RT-PCR分析细胞周期和细胞凋亡

细胞增殖

细胞形态学

hnRNP K和真核细胞翻译起始因子4E（Eukaryotic translation initiation factor-4E

eIF4E）mRNA表达变化。结果:克隆载体序列正确。与空载对照组K562细胞相比

hnRNP K干扰组G2/M期细胞比例明显增多（P<0.05）

而细胞凋亡则没有显著变化（P>0.05）

细胞增殖能力降低

双核和多核细胞明显增多（P<0.05）

hnRNP K和eIF4E mRNA表达较空载对照组分别降低了55.22%和55.27%。结论:hnRNP K基因沉默能使K562细胞周期阻滞于G2/M期

对细胞凋亡没有显著影响

降低细胞增殖能力

下调eIF4E mRNA表达

使双核和多核细胞增多

提示该基因可能与细胞分裂有关。

Abstract

Objective:To explore the effect of heterogeneous nuclear ribonucleoprotein K(hnRNP K)short hairpin RNA(shRNA)on K562 cells. Methods:hnRNP K shRNA retrovirus was packaged with pSUPER retro hnRNP K shRNA after its identification by enzyme digestion and sequencing analysis

then FCM

Wright staining

MTT assay

RT-PCR were used to analyze its effects on cell cycle and apoptosis

proliferation

hnRNP K and eukaryotic translation initiation factor-4E(eIF4E) mRNA of K562 cells respectively. Results:The sequence of the inserted fragment in pSUPER retro hnRNP K shRNA was correct. Compared with the control K562 cells

hnRNP K shRNA treated group showed significantly more cells at G2/M phase(P<0.05)

growth inhibition and more cells with ≥2 nucleus(P<0.05) as well as 55.22% and 55.27% decrease of hnRNP K and eIF4E mRNA respectively. Conclusion:hnRNP K interference blocks K562 cells at G2/M phase

inhibits cell growth

and down-regulates both eIF4E and hnRNP K mRNA expression

increases cells with≥2 nucleus

suggesting that it might play a role during cell division.

关键词

RNA结合蛋白KshRNA逆转录病毒载体真核细胞翻译起始因子4E

Keywords

Heterogeneous nuclear ribonucleoprotein KShort hairpin RNARetroviral vectorEukaryotic translation initiation factor-4E

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