贝伐单抗对体外培养的人视网膜色素上皮细胞增殖及纤维化的影响
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Effects of Bevacizumab on the proliferation and epithelial-mesenchymal transition in human retinal pigment epithelial cells in vitro
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    摘要:

    目的:观察贝伐单抗对体外培养的人视网膜色素上皮细胞ARPE-19增殖及钙黏连蛋白(E-cadherin)和纤维连接蛋白(fibronectin)表达变化的作用,探讨贝伐单抗对ARPE-19增殖及纤维化的影响。

    方法:用不同浓度(0、0.625、1.25、2.5、5.0mg/mL)的贝伐单抗干预体外培养人视网膜色素上皮细胞ARPE-19,采用CCK-8法分别于24、48、72h检测细胞活性; 流式细胞仪检测细胞周期; 应用免疫蛋白印迹法(Western blotting)及逆转录聚合酶链反应(RT-PCR)检测ARPE-19中E-cadherin和fibronectin的蛋白及mRNA的表达变化。

    结果:浓度为2.5、5.0mg/mL贝伐单抗能有效抑制ARPE-19细胞的增殖及细胞周期,差异有统计学意义(P<0.05)。2.5、5.0mg/mL贝伐单抗能抑制E-cadherin基因,促进fibronectin基因的转录及表达,差异有统计学意义(P<0.05)。

    结论:高浓度的贝伐单抗能够抑制ARPE-19细胞的增殖,下调纤维化相关因子E-cadherin,同时上调fibronectin的表达,提示高浓度的贝伐单抗可以引起ARPE-19细胞纤维化。

    Abstract:

    AIM: To investigate the effects of Bevacizumab on the proliferation and the expression of E-Cadherin and fibronectin in human retinal pigment epithelial cell(ARPE-19)in vitro.

    METHODS: Different concentrations(0, 0.625, 1.25, 2.5, 5.0mg/mL)of bevacizumab were exposed to ARPE-19 cells, then cell viability was analyzed by CCK-8, cell cycle was determined by flow cytometry, and the expression of E-Cadherin and fibornectin was detected by Western blot and RT-PCR.

    RESULTS: The concentration as 2.5mg/mL or 5.0mg/mL of bevacizumab was shown to effectively suppress the proliferation and cell cycle of ARPE-19 cell(P<0.05). In addition, 2.5mg/mL or 5.0mg/mL of bevacizumab could downregulate the expression of E-cadherin and promote the transcription of fibronection gene(P<0.05).

    CONCLUSION: High concentration of bevacizumab was able to inhibit ARPE-19 proliferation, downregulate E-Cadherin expression and promote fibronectin expression, indicating epithelial-mesenchymal transition induced by bevacizumab in ARPE-19 cell.

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李婵,任伟.贝伐单抗对体外培养的人视网膜色素上皮细胞增殖及纤维化的影响.国际眼科杂志, 2016,16(8):1449-1452.

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  • 收稿日期:2016-04-26
  • 最后修改日期:2016-07-05
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  • 在线发布日期: 2016-07-26
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