CTGF siRNA转染对体外培养人晶状体上皮细胞株B3转分化的抑制作用
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福建省卫生系统中青年骨干人才培养项目(2014-ZQN-ZD-16)


Transfection of CTGF siRNA inhibits transdifferentiation in human lens epithelium cell line B3 in vitro
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Young and Middle Aged Key Personnel Training Project of Fujian Provincial Health and Family Planning Commission(2014-ZQN-ZD-16)

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    摘要:

    目的:研究脂质体介导CTGF siRNA 转染对人晶状体上皮细胞(HLEC)株B3 CTGF和α-SMA表达的影响。

    方法:5'-异硫氰酸荧光素标记的CTGF siRNA与脂质体混合,并转染HLECs,通过荧光转染评估转染率。我们用CCK-8来评价转染组和对照组的细胞活力并使用实时定量RT-PCR,细胞免疫化学和Western blot来分析CTGF和α-SMA在转染后的表达改变。

    结果:脂质体介导的CTGF siRNA转染呈现出高效的转染率。转染率在24h高达95%。CTGF siRNA 72h转染能有效抑制HLECs的增殖。CTGF siRNA转染24h后CTGF和α-SMA的表达量都显著下降,而对照组则无类似效应。

    结论:CTGF siRNA 能够有效降低CTGF和α-SMA的表达。

    Abstract:

    AIM: To investigate the expression of connective tissue growth factor(CTGF)and α-SMA in human lens epithelium cell(HLEC)line B3 after transfection by liposome-coated siRNA targeting CTGF.

    METHODS: HLECs were transfected with small interfering RNA(siRNA)targeting CTGF, labeled with 5'- fluorescein isothiocyanate(5'-FITC)and coated with lipofectamine. The transfection ratio was evaluated via fluorescence intensity. Cell counting kit-8(CCK-8)assay was performed to assess cytoviability of both non-transfected and transfected HLECs. Quantitative RT- PCR, cell immunochemistry and Western blot analysis were conducted to detect the expression changes of CTGF and α-SMA after transfection.

    RESULTS: A highly effective transfection ratio was observed in siRNA co-transfected with lipofectamine. The transfection ratio reached 95% at 24h. The proliferation of HLECs was inhibited by siRNA after 72h transfection. The expression of CTGF and α-SMA significantly decreased in HLECs after transfected by CTGF siRNA for 24h. This effect was not found in negative control siRNA.

    CONCLUSIONS: SiRNA targeting CTGF decreased CTGF and α-SMA expression in HLECs, which is a potential therapeutic strategy for posterior capsular opacification.

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庄华,郑凝旋,巫婧,等. CTGF siRNA转染对体外培养人晶状体上皮细胞株B3转分化的抑制作用.国际眼科杂志, 2017,17(8):1387-1393.

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  • 收稿日期:2017-01-08
  • 最后修改日期:2017-06-21
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  • 在线发布日期: 2017-07-24
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