[关键词]
[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。
[Key word]
[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.
[中图分类号]
[基金项目]
福建省卫生系统中青年骨干人才培养项目(2014-ZQN-ZD-16)
