Hippo通路在常染色体显性多囊肾病中的作用  

Role of Hippo pathway in autosomal dominant polycystic kidney disease

作　　者：贺靓靓[1] 胡文娟[1] 梅长林[1] 胡惠民[1] 付丽丽[1] 

机构地区：[1]第二军医大学附属长征医院肾内科,上海200003

出　　处：《中华肾脏病杂志》2015年第3期227-232,共6页Chinese Journal of Nephrology

摘　　要：目的 探讨Hippo通路分子在常染色体显性多囊肾病（ADPKD）发病机制中的作用,寻找可能的药物治疗靶点.方法 采用免疫荧光染色、Western印迹和实时定量PCR技术检测Han：SPRD大鼠杂合型和ADPKD患者肾组织Hippo通路分子分布、表达量以及磷酸化水平的差异.小干扰RNA特异性抑制囊肿衬里上皮细胞（WT9-12） YAP （Yes kinaseassociated protein）、TAZ（transcriptional coactivator with PDZ binding motif）和LATS1（large tumor suppressor kinase1）的表达后观察对细胞增殖、凋亡和细胞周期的影响.结果 与野生型大鼠相比,Han：SPRD杂合型大鼠囊肿衬里上皮细胞LATS1表达降低;YAP表达量及去磷酸化活化水平增加;TAZ表达与分布无明显改变.ADPKD患者肾组织中,Hippo通路分子MST1/2（macrophage stimulating1/2）、LATS1 mRNA表达显著低于正常对照（P＜0.05）,而YAP mRNA表达水平显著高于正常对照（P＜0.05）.抑制WT9-12细胞LATS1表达,能促进细胞增殖和分裂;下调YAP表达阻滞细胞于分裂间期,抑制增殖;下调TAZ表达对细胞增殖和周期无显著影响.结论 ADPKD中Hippo通路效应因子YAP去磷酸化活性增强可能是导致疾病发生、发展的重要原因之一.体外实验证实下调YAP表达可抑制肾囊肿衬里上皮细胞分裂增殖,提示YAP的表达和活性是潜在的多囊肾病治疗靶点.Objective To explore the role of Hippo pathway in the pathogenesis of autosomal dominant polycystic kidney disease （ADPKD）,and find potential targets for drug therapy.Methods By means of immunofluorescence staining,Western blotting,Real-time PCR,the differences of sublocalization,expression and phosphorylation level about Hippo pathway molecules in Han：SPRD （cy/＋） and ADPKD patients compared with the control were observed.Knockdown Yes kinaseassociated protein （YAP）,transcriptional coactivator with PDZ binding motif （TAZ） and large tumor suppressor kinase1 （LATS1） in cystic lining epithelium cell line WT9-12 were took by siRNA interference,and then their effects on cell proliferation,apoptosis and cell cycle were assessed.Results In cystic lining epithelium of Han：SPRD（cy/＋）,decreased expression of LATS1 and increased expression of YAP were found compared with the control,and the immunofluorescence of YAP was distributed both in cytoplasm and nucleus,while distribution and expression level of TAZ were without significant variance.Abnormal mRNA expressions of Hippo pathway components in ADPKD patients were found （P ＜ 0.05）.Down-regulation of LATS1 in WT9-12 cells could prohibit phosphorylation of YAP,and prompted proliferation and cell division.Knockdown YAP in WT9-12 cells could inhibited cell proliferation by arresting cell cycle in G0/G1 phase,but down-regulating TAZ showed no significant differences in proliferation and cell cycle.Conclusions Altered Hippo signaling exists in ADPKD,and YAP activation may be one leading cause of autosomal dominant polycystic kidney disease onset.In vitro,knockdown YAP in WT9-12 cells can inhibit cell proliferation by arresting cell cycle and depressing cell division,suggesting the expression level and activity of YAP are potential targets for ADPKD treatment.

关 键 词：多囊肾 常染色体显性 细胞增殖 细胞周期 Hippo信号通路 YAP LATS1 

分 类 号：R692.1[医药卫生—泌尿科学]