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[促红细胞生成素基因对肺腺癌细胞系A549生长和凋亡的影响。]

[Effects of EPO Gene on Growth and Apoptosis of Lung Adenocarcinoma Cell Line A549.].

作者信息

Chen Bo, Zhao Weihong, Wu Jianqing

机构信息

Department of Geriatrics, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.

出版信息

Zhongguo Fei Ai Za Zhi. 2009 Sep 20;12(9):956-60. doi: 10.3779/j.issn.1009-3419.2009.09.003.

Abstract

BACKGROUND

Published data on the association between erythropoietin (EPO) and cancer cell are inconclusive. The aim of this study is to investigate the effect of erythropoietin (EPO) on the growth and survival of lung adenocarcinoma cell line A549.

METHODS

The recombinant plasmid pcDNA3.1(-)-hEPO was constructed and transfected into A549 cells by liposome protoco1. The Levels of EPO in culture supernatant were detected by ELISA. Effects of EPO gene on growth and survival of the transfected cells were evaluated by MTT assay and flow cytometry (FCM ). Levels of vascular endothelial growth factor (VEGF) were also evaluated by ELISA.

RESULTS

The recombinant eukaryotic expression vector pcDNA3.1(-)-hEPO was successfully constructed. The growth of cells in hEPO transfected cells was significantly inhibited after transfection (P<0.01). More cells were blocked in S phase in hEPO transfected group compared with control group (P<0.05), and the apoptotic rate were also significantly higher than those of their controls (P<0.01). Levels of VEGF in hEPO transfected cells were significantly lower than controls (P<0.01).

CONCLUSIONS

Exogenous EPO gene expression in A549 cells can induce cell growth inhibition and apoptosis of A549 cells, and expression of VEGF can also be inhibited.

摘要

背景

关于促红细胞生成素(EPO)与癌细胞之间关联的已发表数据尚无定论。本研究旨在探讨促红细胞生成素(EPO)对肺腺癌细胞系A549生长和存活的影响。

方法

构建重组质粒pcDNA3.1(-)-hEPO,并通过脂质体protocol将其转染至A549细胞中。采用酶联免疫吸附测定法(ELISA)检测培养上清液中EPO的水平。通过MTT法和流式细胞术(FCM)评估EPO基因对转染细胞生长和存活的影响。同时也采用ELISA评估血管内皮生长因子(VEGF)的水平。

结果

成功构建了重组真核表达载体pcDNA3.1(-)-hEPO。转染后,hEPO转染细胞的生长受到显著抑制(P<0.01)。与对照组相比,hEPO转染组更多细胞停滞于S期(P<0.05),且凋亡率也显著高于对照组(P<0.01)。hEPO转染细胞中VEGF的水平显著低于对照组(P<0.01)。

结论

A549细胞中外源性EPO基因表达可诱导A549细胞生长抑制和凋亡,同时也可抑制VEGF的表达。

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