中文摘要：

目的：观察常染色体显性多囊肾病（ADPKD）患者及动物模型中泛素连接酶β-TrCP 的表达，初步探讨人囊肿衬里上皮 OX161细胞中β-TrCP 的作用和调控机制。方法在 OX161细胞内转染β-TrCP 的 siRNA，采用蛋白免疫印迹法检测增殖细胞核抗原（PCNA）的表达，MTT 比色法检测吸光度；在 OX161细胞中抑制 JAK2／STAT3通路或转染 STAT3质粒，采用蛋白免疫印迹法检测 JAK2、p-JAK2、STAT3、p-STAT3和β-TrCP 的表达。收集ADPKD 行肾脏切除术患者的肾脏组织和肾旁距癌组织5 cm 以上的正常肾脏组织（正常对照组）、雄性 Han：SPRD （cy／＋）大鼠（ADPKD 模型）和野生型 Han：SPRD（＋／＋）大鼠肾组织（对照），均采用蛋白免疫印迹法检测β-TrCP的表达。收集 Pkd1flox／flox；tamoxifen-Cre＋小鼠（ADPKD 模型）和 Pkd1flox／flox；tamoxifen-Cre-小鼠（对照）肾组织，采用免疫组织化学法检测β-TrCP 的表达 和 亚 定 位。结果与 UCL93细胞相比，OX161细 胞 中 JAK2／STAT3通路明显活化，β-TrCP 表达量增加；β-TrCP 敲减后细胞增殖减慢（P ＜0．05）；WP1006抑制 JAK2／STAT 3通路后，β-TrCP 表达量减低，转染 STAT 3质粒后，β-TrCP 的表达量增加。与正常对照组、野生型 Han：SPRD（＋／＋）大鼠及 Pkd1flox／flox；tamoxifen-Cre-小鼠比较，ADPKD 患者肾组织、Han：SPRD（Cy／＋）大鼠肾组织及 Pkd1flox／flox；tamoxifen-Cre＋小鼠肾组织中β-TrCP 表达量明显增加。结论在人囊肿衬里上皮 OX161细胞株中β-TrCP表达量增多，且受 JAK2／STAT3通 路 调 控；在 ADPKD 患者及其模型鼠肾脏组织中β-TrCP 表达量也增加。β-TrCP可能参与促进囊肿的发生和发展。

英文摘要：

Objective To observe the expression of ubiquitin ligaseβ-TrCP in animal model and patients with autosomal dominant polycystic kidney disease（ADPKD）,and to preliminarily inves-tigate the function and regulatory mechanism ofβ-TrCP in human OX161 cells.Methods OX161 cells were transfected with β-TrCP siRNA.The proliferation of OX161 cells was measured by＆nbsp;MTT assay,and the expression of PCNA was detected by Western blot.Furthermore,OX161 cells were treated with JAK2/STAT3 inhibitor WP1006 or transfected with STAT3 expression plas-mids.The expression of JAK2,p-JAK2,STAT3,p-STAT3 andβ-TrCP was examined by Western blot.Moreover,kidney tissues and normal tissues 5 cm from cancer tissues of ADPKD patients undergoing nephrectomy and kidney tissues of male Han：SPRD（cy/＋）rats and Pkd1flox/flox：tamoxifen-Cre＋ mice and wild type Han：SPRD（＋/＋）rats and Pkd1flox/flox：tamoxifen-Cre-mice were collected to determine the expression and localization of β-TrCP.Results Compared with UCL93 cells,JAK2/STAT3 pathway was activated andβ-TrCP expression was increased in OX161 cells.The proliferation of OX161 cells was suppressed after knockdown of β-TrCP.The expression of β-TrCP decreased after treatment with WP1006,but increased after transfection with STAT3 plasmids.Compared with normal kidney tissues from ADPKD patients,kidney tis-sues from wild type Han：SPRD（＋/＋）rats and kidney tissues from Pkd1flox/flox：tamoxifen-Cre-mice,the expression of β-TrCP respectively increased in kidney tissues from ADPKD pa-tients,Han：SPRD（cy/＋）rats and Pkd1flox/flox：tamoxifen-Cre＋ mice.Conclusion The ex-pression ofβ-TrCP increases in human OX161 cells,as well as in kidney tissues from ADPKD pa-tients and animal models,suggesting thatβ-TrCP is involved in the occurrence and development of ADPKD.In addition,β-TrCP expression is regulated by JAK2/STAT3 pathway.