Erythropoietin promotes the proliferation of glioblastoma cells through up-regulation of Cyclin D1 in vitro

Wang Wentao; Tang Zhaohua; Huo Gang; Li Ying; Wang Haiquan; Ye Xiaolin; Liu Zili; Tan Ying; Shi Shuang

doi:10.13406/j.cnki.cyxb.000978

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Erythropoietin promotes the proliferation of glioblastoma cells through up-regulation of Cyclin D1 in vitro

更新时间：2024-01-17

- Erythropoietin promotes the proliferation of glioblastoma cells through up-regulation of Cyclin D1 in vitro
- Journal of Chongqing Medical University Vol. 41, Issue 8, Pages: 787-791(2016)
- 作者机构：
  
  重庆医科大学附属第一医院神经外科
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.000978
  CLC： R739.41
- Published：2016，
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[1]王文涛,唐兆华,霍钢,李英,王海全,叶小琳,刘自力,谭赢,石爽.EPO通过上调CyclinD1表达促进胶质母细胞瘤的体外增殖[J].重庆医科大学学报,2016,41(08):787-791.

Wang Wentao, Tang Zhaohua, Huo Gang, et al. Erythropoietin promotes the proliferation of glioblastoma cells through up-regulation of Cyclin D1 in vitro. [J]. Journal of Chongqing Medical University 41(8):787-791(2016)
[1]王文涛,唐兆华,霍钢,李英,王海全,叶小琳,刘自力,谭赢,石爽.EPO通过上调CyclinD1表达促进胶质母细胞瘤的体外增殖[J].重庆医科大学学报,2016,41(08):787-791. DOI： 10.13406/j.cnki.cyxb.000978.

Wang Wentao, Tang Zhaohua, Huo Gang, et al. Erythropoietin promotes the proliferation of glioblastoma cells through up-regulation of Cyclin D1 in vitro. [J]. Journal of Chongqing Medical University 41(8):787-791(2016) DOI： 10.13406/j.cnki.cyxb.000978.

摘要

目的:研究促红细胞生成素（erythropoietin

EPO）对胶质母细胞瘤体外增殖的影响

并探讨其机制。方法:体外培养胶质母细胞瘤U87细胞株

实验分为对照组、EPO处理组及EPO+细胞周期素D1（Cyclin D1）拮抗剂组。对照组:用等体积PBS替代EPO;EPO处理组:在U87细胞的培养基中加入EPO（0.1 U/ml）处理7d;EPO+cyclin D1拮抗剂组:U87细胞的培养基中同时加入EPO（0.1 U/ml）及Cyclin D1拮抗剂（5μmol/L）共处理7 d。用Cell Counting Kit-8（CCK-8）法测细胞增殖情况

细胞倍增时间检测各组细胞增殖速度

RT-PCR、Western blot和细胞免疫荧光法（immunofluorescence

IF）测定细胞周期关键蛋白Cyclin D1的m RNA和蛋白表达变化

流式细胞仪检测各组细胞周期。结果:与对照组相比

EPO处理组中U87细胞生长速度明显增快（CCK8:第3天P=0.014

第5天P=0.029

倍增时间:P=0.012）

细胞中Cyclin D1的m RNA及蛋白表达水平也明显升高

差异有统计学意义（m RNA:P=0.001

蛋白:P=0.002

荧光定量:P=0.005）

流式细胞仪检测发现U87细胞的增殖指数也明显升高（P=0.000）;与EPO处理组相比

EPO+cyclin D1拮抗剂组细胞Cyclin D1 m RNA及蛋白的表达水平明显降低（m RNA:P=0.009

蛋白:P=0.000

荧光定量:P=0.013）

流式细胞仪检测细胞增殖指数则有显著下降（P=0.000）

同时细胞增殖能力和速度较EPO处理组明显降低（CCK8:第3天P=0.000

第5天P=0.000

倍增时间:P=0.001）。结论:本研究发现EPO可显著促进胶质母细胞瘤的体外增殖

上调细胞周期关键蛋白Cyclin D1的表达

加快细胞增殖周期可能是其重要的分子机制。

Abstract

Objective:To explore the effect of recombinant human erythropoietin(rh EPO)on the proliferation of glioblastoma cells in vitro and its mechanism. Methods:U87 cells were cultured in vitro and divided into three groups:control group treated with equal value of PBS;EPO treatment group cultured with a dose of 0.1 U/ml EPO for seven days;EPO + Cyclin D1 antagonist group treated with EPO(0.1 U/ml)and Cyclin D1 antagonist(5 μmol/L)for seven days. The proliferative ability was measured by cell counting Kit-8 assay and double time;Cyclin D1 expression was detected by RT-PCR

Western blot and cell immunofluorescence

respectively. The cell cycle was analyzed by flow cytometry. Results:EPO promoted the proliferation of U87 cells and the m RNA and protein levels of Cyclin D1. Cyclin D1 antagonist inhibited the m RNA and protein levels of Cyclin D1 induced by EPO;U87 cells were blockaded at G1 stage

and the proliferation of ability was significantly reduced. Conclusion:EPO may significantly promote the proliferation of U87 cells through upregulation the expression of Cyclin D1;its molecular mechanism may relate with accelerated cell proliferation cycle.

关键词

促红细胞生成素胶质母细胞瘤Cyclin D1细胞周期增殖Cyclin D1拮抗剂

Keywords

erythropoietinglioblastomaCyclin D1cell cycleproliferationCyclin D1 antagonist

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