Cai Yumei, Yang Weiqun, Pan Mingmeng, Wang Chaoyang, Wu Wenyi, Zhu Shize
Department of Pathology, Quanzhou Medical College Department of Dermatology Department of Plastic Surgery, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, Fujian, China.
Medicine (Baltimore). 2018 Sep;97(37):e12167. doi: 10.1097/MD.0000000000012167.
To study the effect of knocking down wingless-related MMTV integration site 2 (Wnt2) expression by RNAi on the growth and signaling pathways of ex vitro-cultured keloid fibroblasts (KFB).Human KFB were isolated from 10 keloid patient specimens. The KFB cells were then transfected with 4 pairs of small interfering RNA (siRNA) targeting human Wnt2, respectively. Reverse transcriptase-polymerase chain reaction and Western blot analysis were conducted to verify the knock down of Wnt2, and the expression of β-catenin glycogen synthase kinase-3β (GSK-3β) and cyclin D1 were examined.siRNA Wnt2 transfection (siWnt2) resulted in the significant inhibition of Wnt2 expression at both the mRNA and protein levels. The expression of β-catenin, GSK-3β, p-GSK-3β, and cyclin D1 at the protein level also decreased in siWnt2 cells. siWnt2 resulted in a substantially slower growth and significant delay in cell doubling time of the KFB cells compared with control groups. Further, the siRNA knock down of GSK-3β and β-catenin resulted in slower proliferation rates, respectively.Wnt2 siRNA has an inhibitive effect on keloid fibroblast proliferation, which may be a potential therapeutic approach for keloid and other human fibrotic diseases.
研究RNA干扰敲低无翅相关MMTV整合位点2（Wnt2）表达对体外培养的瘢痕疙瘩成纤维细胞（KFB）生长及信号通路的影响。从10例瘢痕疙瘩患者标本中分离出人KFB。然后分别用4对靶向人Wnt2的小干扰RNA（siRNA）转染KFB细胞。进行逆转录聚合酶链反应和蛋白质印迹分析以验证Wnt2的敲低，并检测β-连环蛋白、糖原合酶激酶-3β（GSK-3β）和细胞周期蛋白D1的表达。siRNA Wnt2转染（siWnt2）导致Wnt2在mRNA和蛋白质水平均受到显著抑制。siWnt2细胞中β-连环蛋白、GSK-3β、p-GSK-3β和细胞周期蛋白D1的蛋白质水平表达也降低。与对照组相比，siWnt2导致KFB细胞生长明显减慢，细胞倍增时间显著延迟。此外，GSK-3β和β-连环蛋白的siRNA敲低分别导致增殖速率减慢。Wnt2 siRNA对瘢痕疙瘩成纤维细胞增殖有抑制作用，这可能是瘢痕疙瘩及其他人类纤维化疾病的一种潜在治疗方法。
