Huang Wan-Rong, Fan Xing-Xing, Tang Xin
Tianjin Eye Hospital, Tianjin Medical University, Tianjin, China.
Mol Vis. 2011;17:2349-55. Epub 2011 Aug 31.
We investigated the effect of epidermal growth factor receptor (EGFR) siRNA on human lens epithelium (HLE) cells and the development of posterior capsular opacity (PCO).
We designed EGFR siRNA and used it to knockdown the expression of EGFR in HLE cells. Cell proliferation was examined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), cell growth curve assay and cell cycle analysis. Next, we selected an adaptable concentration of recombinant epidermal growth factor (EGF) for stimulating the growth of HLE cells to further test the suppressive effect of siRNA. At last, we established the model of PCO in rats to further investigate whether knocking down EGFR would prevent the progression of PCO in vivo.
The cell proliferation of EGFR siRNA group was apparently inhibited no matter in short or long term and cell cycle was arrested in G(1) phase. Over expression EGF cannot rescue the inhibition of EGFR siRNA on HLE cells and the proliferation activity in HLE cells greatly decreased when EGF-EGFR signal pathway blockaded. In vivo experiments, the extent of PCO of EGFR siRNA group is much lower than the control group.
Our results demonstrate that EGFR siRNA can effectively inhibit the progression of PCO. Thus, siRNA targeting EGFR may provide a totally new way for preventing PCO or even cataract.
我们研究了表皮生长因子受体（EGFR）小干扰RNA（siRNA）对人晶状体上皮（HLE）细胞及后囊膜混浊（PCO）发展的影响。
我们设计了EGFR siRNA，并用于敲低HLE细胞中EGFR的表达。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）、细胞生长曲线分析和细胞周期分析来检测细胞增殖。接下来，我们选择合适浓度的重组表皮生长因子（EGF）来刺激HLE细胞生长，以进一步测试siRNA的抑制作用。最后，我们建立大鼠PCO模型，以进一步研究敲低EGFR是否能在体内阻止PCO的进展。
无论短期还是长期，EGFR siRNA组的细胞增殖均明显受到抑制，细胞周期停滞于G(1)期。EGF过表达不能挽救EGFR siRNA对HLE细胞的抑制作用，当EGF-EGFR信号通路被阻断时，HLE细胞的增殖活性大大降低。在体内实验中，EGFR siRNA组的PCO程度远低于对照组。
我们的结果表明，EGFR siRNA可有效抑制PCO的进展。因此，靶向EGFR的siRNA可能为预防PCO甚至白内障提供一种全新的方法。
