Effect of ILK SiRNA on the proliferation and apoptosis of human Tenon fibroblasts induced by transforming growth factor-β₂

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 group: HTFs+5μg/L TGF-β2; H+T+NC SiRNA group: HTFs+5μg/L TGF-β2+50nmol/L negtive SiRNA; H+T+ILK SiRNA group: HTFs+5μg/L TGF-β2+50nmol/L ILK SiRNA. The ILK SiRNA were transfected into HTFs by lipofectamine 2000, then the cells were stimulated with 5μg/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-β2 treated and control groups, and TGF-β2 up-regulated the expression of ILK, ILK SiRNA inhibited the protein expression of ILK(P2 treated group increased, and in ILK SiRNA group the cell proliferation rate was suppressed after exposured to ILK SiRNA for 48h(P2 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(PCONCLUSION: The ILK SiRNA attenuates the abnormal proliferation of HTFs induced by TGF-β2, thereby enhancing the apoptosis of HTFs.

Keywords

• Integrin-linked kinase
• fibroblasts
• transforming growth factor-β₂
• proliferation
• apoptosis
• apoptosis