Effect of ILK SiRNA on the proliferation and apoptosis of human Tenon fibroblasts induced by transforming growth factor-β2 / 国际眼科杂志(Guoji Yanke Zazhi)

ABSTRACT

·AIM: To investigate the role of small interference RNA interference targeted Integrin-linked kinase (ILK SiRNA) on the proliferation and apoptosis of human Tenon fibroblasts (HTFs) induced by transforming growth factor-β2(TGF-β2). · METHODS: The HTFs were identified by immunofluorescence analysis with Vimentin and keratin. HTFs with no other addiction was as normal control;H+T groupHTFs+5μ g/L TGF-β2;H+T+NC SiRNA groupHTFs+5μ g/L TGF-β2+50nmol/L negtive SiRNA; H+T+ILK SiRNA groupHTFs+5μ g/L TGF-β2+50nmol/L ILK SiRNA. The ILK SiRNA were transfected into HTFs by lipofectamine 2000, then the cells were stimulated with 5μ g/L TGF - β2. The protein expression of ILK were analyzed by Western Blot. The proliferation levels of HTFs were analyzed by CCK-8, the apoptosis of HTFs were analyzed by Hoechst 33342/PI double staining. ·RESULTS: The protein ILK were expressed in both TGF-β2treated and control groups, and TGF- β2up-regulated the expression of ILK, ILK SiRNA inhibited the protein expression of ILK(P< 0. 05). CCK- 8 analysis showed that compared with the normal control group,the cell proliferation rate of HTFs in TGF-β2treated group increased, and in ILK SiRNA group the cell proliferation rate was suppressed after exposured to ILK SiRNA for 48h (P<0.05). Hoechst 33342/PI double staining showed that there was no change on the apoptosis of TGF - β2 stimulated group (P>0.05), compared with the normal control group, however in the ILK SiRNA group, we found lots of apoptosis cells and a few of necrotic cells (P<0.05). ·CONCLUSION: The ILK SiRNA attenuates the abnormal proliferation of HTFs induced by TGF - β2, thereby enhancing the apoptosis of HTFs.