RNA干扰敲低GSK-3β对瘢痕疙瘩形成影响的体外研究  

Effects of GSK-3β knockdown by RNA interference on formation of keloid in vitro

作　　者：蔡玉梅 朱世泽[2] 杨维群 潘明孟[2] 王朝阳[2] 吴文艺[2] 

机构地区：[1]泉州医学高等专科学校病理学教研室,福建泉州362000 [2]福建医科大学附属第二医院整形外科,福建泉州362000

出　　处：《中国病理生理杂志》2017年第1期154-160,共7页Chinese Journal of Pathophysiology

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)

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。AIM：To study the suppressive effect of glycogen synthase kinase-3β（GSK-3β）knockdown by RNA interference on the formation of keloid. METHODS：Human keloid fibroblasts（KFB）in vitro were transfected with3 pairs of specific GSK-3β small interfering RNA（siRNA）. The best siRNA to inhibit the GSK-3β expression in human KFB was screen by RT-PCR and Western blot. The expression of GSK-3β and related proteins at m RNA and protein levels in the KFB was determined by RT-PCR and Western blot. RESULTS：The GSK-3β siRNA1434 remarkably inhibited the expression of GSK-3β at m RNA and proteins levels in the human KFB. After transfection with GSK-3β siRNA,the protein levels of β-catenin,p-GSK-3β,Wnt2 and cyclin D1 were all decreased. KFB growth became slow. With the extension of time,the inhibition of cell growth increased,and the cell doubling time was significantly delayed. CONCLUSION：siRNA targeting GSK-3β efficiently knocks down the expression of GSK-3β in the human KFB,and inhibits the activation of Wnt signaling pathway,thus inhibiting the growth of keloid. GSK-3β may be a potential therapeutic target for keloid.

关 键 词：糖原合成酶激酶3Β 瘢痕疙瘩 成纤维细胞 RNA干扰 

分 类 号：R363[医药卫生—病理学] R751[医药卫生—基础医学]