Effect of ILK SiRNA on the proliferation and apoptosis of human Tenon fibroblasts induced by transforming growth factor-β<sub>2</sub>
Journal Title: Guoji Yanke Zazhi - Year 2018, Vol 18, Issue 4
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-β<sub>2</sub><sub> </sub>(TGF-β<sub>2</sub>). <p>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-β<sub>2</sub>; <sub> </sub>H+T+NC SiRNA group: HTFs+5μg/L TGF-β<sub>2</sub>+50nmol/L negtive SiRNA; H+T+ILK SiRNA group: HTFs+5μg/L TGF-β<sub>2</sub>+50nmol/L ILK SiRNA. The ILK SiRNA were transfected into HTFs by lipofectamine 2000, then the cells were stimulated with 5μg/L TGF-β<sub>2</sub>. 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. <p>RESULTS: The protein ILK were expressed in both TGF-β<sub>2</sub> treated and control groups, and TGF-β<sub>2</sub> up-regulated the expression of ILK, ILK SiRNA inhibited the protein expression of ILK(<i>P</i><0.05). CCK-8 analysis showed that compared with the normal control group, the cell proliferation rate of HTFs in TGF-β<sub>2</sub> treated group increased, and in ILK SiRNA group the cell proliferation rate was suppressed after exposured to ILK SiRNA for 48h(<i>P</i><0.05). Hoechst 33342/PI double staining showed that there was no change on the apoptosis of TGF-β<sub>2</sub> stimulated group(<i>P</i>>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(<i>P</i><0.05). <p>CONCLUSION: The ILK SiRNA attenuates the abnormal proliferation of HTFs induced by TGF-β2, thereby enhancing the apoptosis of HTFs.
Authors and Affiliations
Qian - Yan Kang
Keywords
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- EP ID EP656040
- DOI 10.3980/j.issn.1672-5123.2018.4.06
- Views 119
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