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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    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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Hua Zhuang, Ning-Xuan Zheng, Jing Wu, et al. Transfection of CTGF siRNA inhibits transdifferentiation in human lens epithelium cell line B3 in vitro. Guoji Yanke Zazhi( Int Eye Sci) 2017;17(8):1387-1393

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Publication History
  • Received:January 08,2017
  • Revised:June 21,2017
  • Adopted:
  • Online: July 24,2017
  • Published: