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[Stat3激活降低对人胃癌细胞系5-氟尿嘧啶耐药性的影响]

[The impact of decreased Stat3 activation on 5-fluorouracil resistance of human gastric cancer cell line].

作者信息

Yu Li-fen, Chen Ying, Qiao Min-min, Zhang Yong-ping, Wu Yun-lin

机构信息

Department of Gastroenterology, Ruijin Hospital of Shanghai Second Medical University, Shanghai, 200025 China.

出版信息

Zhonghua Nei Ke Za Zhi. 2004 Dec;43(12):903-6.

Abstract

OBJECTIVE

To investigate the relationship between different activation of Stat3 signaling and the drug resistance mechanisms in two human gastric cancer cell lines, 5-fluorouracil (5-FU) resistant cell line and its parental cell line.

METHODS

Electrophoretic mobility shift assay and Western blot were used to detected Stat3 DNA-binding activity and the expression of phospho-Stat3 protein in 5-FU resistant cell line SGC7901/R and its parental cell line SGC7901, respectively. The mRNA expression of Stat3 and vascular endothelial growth factor (VEGF) were analysed with semi-quantitative RT-PCR. The expressive intensity of VEGF protein was measured by immunocytochemistry.

RESULTS

The constitutive activation of Stat3 and the expression of phospho-Stat3 protein were different in two human gastric cancer cell lines. Compared with the parental cell line SGC7901, the Stat3-DNA binding activity and the expressive intensity of phospho-Stat3 protein were lower in the drug-resistant cell line SGC7901/R. The expression level of Stat3 mRNA was also decreased in drug resistant cell line, so did VEGF mRNA and its encoded protein.

CONCLUSIONS

The decreased Stat3 activation in 5-FU resistant human gastric cancer cell line SGC7901/R is related to the drug resistance mechanisms and may be correlated with the lower VEGF expression.

摘要

目的

研究两种人胃癌细胞系(5-氟尿嘧啶(5-FU)耐药细胞系及其亲本细胞系)中Stat3信号通路的不同激活状态与耐药机制之间的关系。

方法

分别采用电泳迁移率变动分析和蛋白质免疫印迹法检测5-FU耐药细胞系SGC7901/R及其亲本细胞系SGC7901中Stat3的DNA结合活性和磷酸化Stat3蛋白的表达。采用半定量逆转录-聚合酶链反应(RT-PCR)分析Stat3和血管内皮生长因子(VEGF)的mRNA表达。通过免疫细胞化学法检测VEGF蛋白的表达强度。

结果

两种人胃癌细胞系中Stat3的组成性激活及磷酸化Stat3蛋白的表达存在差异。与亲本细胞系SGC7901相比,耐药细胞系SGC7901/R中Stat3的DNA结合活性及磷酸化Stat3蛋白的表达强度较低。耐药细胞系中Stat3 mRNA的表达水平也降低,VEGF mRNA及其编码蛋白的表达水平同样降低。

结论

5-FU耐药人胃癌细胞系SGC7901/R中Stat3激活的降低与耐药机制有关,可能与VEGF表达降低有关。

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