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可溶性血管内皮生长因子受体-1基因转染人多发性骨髓瘤细胞的体外研究

[In vitro study of human multiple myeloma cell transfected with soluble vascular endothelial growth factor receptor-1 gene].

作者信息

Zheng Dong, Li Juan, Liu Jun-Ru, Zhou Zhen-Hai, Gu Jing-Li

机构信息

Department of Hematology, The First Affiliated Hospital of Sun Yat-sen University,Guangzhou, Guangdong 510080, People's Republic of China.

出版信息

Chin J Cancer. 2010 Jan;29(1):65-8.

Abstract

BACKGROUND AND OBJECTIVE

Vascular endothelial growth factor (VEGF) is the most important angiogenic factor of multiple myeloma (MM). This study was to investigate the effect of transfection of human soluble vascular endothelial growth factor receptor-1 (sFlt-1) gene on the proliferation of human MM cell line RPMI8226.

METHODS

The recombinant plasmid pcDNA3-sFlt-1 was constructed and transfected into RPMI8226 cells. The expression of sFlt-1 was identified by reverse transcription-polymerase chain reaction (RT-PCR) and ELISA. The effects of sFlt-1 protein on the proliferation and VEGF expression of RPMI8226 cells were investigated by MTT assay and ELISA, respectively.

RESULTS

The recombinant plasmid pcDNA3-sFlt-1 was successfully transfected into RPMI8226 cells. sFlt-1 protein expression was identified by ELISA, which inhibited the proliferation of RPMI8226 cells and reduced VEGF concentration in the culture supernatant.

CONCLUSION

RPMI8226 cells can express sFlt-1 protein with high biological activity when transfected with the sFlt-1 gene, which inhibits the proliferation of RPMI8226 cells.

摘要

背景与目的

血管内皮生长因子(VEGF)是多发性骨髓瘤(MM)最重要的血管生成因子。本研究旨在探讨转染人可溶性血管内皮生长因子受体-1(sFlt-1)基因对人MM细胞系RPMI8226增殖的影响。

方法

构建重组质粒pcDNA3-sFlt-1并转染至RPMI8226细胞。通过逆转录聚合酶链反应(RT-PCR)和酶联免疫吸附测定(ELISA)鉴定sFlt-1的表达。分别采用MTT法和ELISA法研究sFlt-1蛋白对RPMI8226细胞增殖和VEGF表达的影响。

结果

重组质粒pcDNA3-sFlt-1成功转染至RPMI8226细胞。通过ELISA鉴定sFlt-1蛋白表达,其抑制RPMI8226细胞增殖并降低培养上清液中VEGF浓度。

结论

RPMI8226细胞转染sFlt-1基因后可表达具有高生物活性的sFlt-1蛋白,抑制RPMI8226细胞增殖。

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