卷曲蛋白受体小干扰RNA对瘢痕疙瘩形成影响的体外研究  

Effects of targeted inhibition of Frizzled receptor with siRNA on the formation of keloid

作　　者：蔡玉梅 朱世泽[2] 杨维群 潘明孟[3] 王朝阳[2] 吴文艺[2] CAI Yu-mei;ZHU Shi-ze;YANG Wei-qun;PAN Ming-meng;WANG Chao-yang;WU Wen-yi(Department of Pathology,Quanzhou Medical College,Quanzhou 362000,Chin)

机构地区：[1]泉州医学高等专科学校病理教研室,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 [3]福建医科大学附属第二医院皮肤科,福建泉州362000

出　　处：《临床皮肤科杂志》2018年第8期496-500,共5页Journal of Clinical Dermatology

基　　金：福建省泉州市科技局重点资助科技项目(2012Z70)资助项目

摘　　要：目的 :探讨RNA干扰技术(RNAi)靶向抑制卷曲蛋白(Frizzled)受体对人瘢痕疙瘩成纤维细胞(KFB)的抑制作用。方法:将针对人Frizzled受体基因设计合成的3对特异性小干扰RNA(si RNA)分别转染体外培养的人KFB,通过实时荧光定量聚合酶链式反应(q PCR)和western blot筛选出干扰人KFB Frizzled受体基因表达的最佳si RNA,进而转染人KFB,并采用q PCR及western blot检测Frizzled受体及相关基因和蛋白表达;采用细胞增殖检测试剂(CCK)-8检测细胞增殖情况。结果:si RNA303具有最佳的Frizzled受体m RNA和蛋白质抑制效率,Frizzled受体si RNA转染后,KFB的Frizzled受体、Wnt2、糖原合酶激酶(GSK)3β和D型细胞周期蛋白(cyclin D)1 m RNA及蛋白表达均下降。p-GSK3β蛋白表达下降,KFB生长受抑制程度增大,增殖减慢,且随着培养时间的延长,细胞倍增时间明显延迟。结论:转染靶向Frizzled受体的si RNA可有效敲低该基因在KFB内的表达,从而抑制成纤维细胞增殖。Objective： To study the suppressive effects of knocking down the expression of Frizzled receptor with small interfering RNA（siRNA） on keloid formation in vitro. Method： Human keloid fibroblasts（KFB） were transfected by 3 pairs of specific siRNA, which were designed and synthesized against the human Frizzled receptor gene. Quantitative real-time polymerase chain reaction（qPCR） and western blot were used to identify the most effective siRNA inhibiting expression of Frizzled receptor in human KFB. Following transfection with the most effective siRNA, expression levels of related genes and proteins were assessed using qPCR and western blot, respectively. Cell proliferation was evaluated using CCK-8 assay. Results： The 303 siRNA displayed the most potent inhibition on both gene and protein expression. Transfection with 303 siRNA decreased ex pression levels of both mRNA and proteins of Frizzled receptor, Wnt2, GSK3β and cyclin D1 in addition to a reduction in expression of p-GSK3β protein. The extent of inhibition on KFB growth positively correlated with the treatment time. Cell doubling time was significantly delayed. Conclusions： Transfection with target siRNA target can knockdown Frizzled receptor gene, leading to inhibition of KFB growth.

关 键 词：卷曲蛋白受体 瘢痕疙瘩 成纤维细胞 RNA干扰 

分 类 号：R619.6[医药卫生—外科学]