RNA干扰敲低GSK-3β对瘢痕疙瘩形成影响的体外研究

发布时间：2018-12-06 16:07

【摘要】：目的:利用RNA干扰技术探讨糖原合成酶激酶3β(GSK-3β)对人瘢痕疙瘩成纤维细胞(keloid fibroblasts,KFB)的抑制效果。方法:将针对人GSK-3β基因设计合成的3对特异性小干扰RNA(siRNA)分别转染体外培养的人KFB,通过RT-PCR和Western blot筛选出干扰人KFB GSK-3β基因表达的最佳siRNA,进而转染人KFB,并用RT-PCR和Western blot检测GSK-3β及相关蛋白的m RNA和蛋白表达。结果:1434序列具有最佳的GSK-3βm RNA和蛋白抑制效率。转染GSK-3βsiRNA后,KFB的β-catenin、细胞周期蛋白D1(cyclin D1)、p-GSK-3β和Wnt2的蛋白水平下降,KFB活力下降,且随着培养时间的延长,细胞生长受抑制程度增大,细胞倍增时间明显延迟。结论:转染靶向GSK-3β的siRNA可有效降低该基因在KFB内的表达,从而抑制了瘢痕疙瘩生长,具有潜在的治疗前景。
[Abstract]:Aim: to investigate the inhibitory effect of glycogen synthase kinase 3 尾 (GSK-3 尾) on human keloid fibroblasts (keloid fibroblasts,KFB) by RNA interference technique. Methods: three pairs of specific small interfering RNA (siRNA) designed and synthesized for human GSK-3 尾 gene were transfected into human KFB, cultured in vitro. The best siRNA, that interfered with the expression of KFB GSK-3 尾 gene was screened by RT-PCR and Western blot, and then transfected into human KFB,. The expression of m RNA and protein of GSK-3 尾 and related proteins was detected by RT-PCR and Western blot. Results: 1434 sequence had the best inhibition efficiency of GSK-3 尾 m RNA and protein. After transfection of GSK-3 尾 siRNA, the levels of cyclin D1 (cyclin D1), p-GSK-3 尾 and Wnt2 in 尾 catenin, cells of KFB decreased, KFB activity decreased, and the inhibition degree of cell growth increased with the extension of culture time. Cell doubling time was significantly delayed. Conclusion: transfection of siRNA targeting GSK-3 尾 can effectively reduce the expression of the gene in KFB and thus inhibit keloid growth.
【作者单位】：泉州医学高等专科学校病理学教研室;福建医科大学附属第二医院整形外科;
【基金】：福建省泉州市科技局重点资助科技项目(No.2012Z70) 福建省医学创新课题(No.2009-CX-21)
【分类号】：R622

【参考文献】