Yes相关蛋白1对胆囊癌细胞增殖及迁移的影响  

Influence of Yes-associated protein-1 on proliferation and metastasis of gallbladder carcinoma

作　　者：何睿[1] 汪洋[1] 黄颖滨 杜军[2] 焦兴元[1] 

机构地区：[1]中山大学附属第一医院器官移植中心,广州510080 [2]中山大学药学院分子生物实验室

出　　处：《中华肝胆外科杂志》2017年第2期95-99,共5页Chinese Journal of Hepatobiliary Surgery

基　　金：国家自然科学基金面上项目（81270531）;广东省科技计划项目（2014A020212485）

摘　　要：目的探讨正常胆囊黏膜上皮、胆囊黏膜单纯性增生、不典型增生及胆囊癌黏膜上皮中Yes相关蛋白1（YAP1）的表达及其在胆囊癌病程发展中的作用和机制。方法应用免疫组化法检测50例正常胆囊、101例慢性胆囊炎／胆囊结石及100例原发性胆囊癌患者的胆囊活检标本石蜡切片中胆囊黏膜上皮细胞类型及YAPl蛋白的表达与分布情况。原代培养胆囊黏膜上皮细胞，应用实时PCR（RT—PCR）法和蛋白印迹检测（Western blot）法比较正常黏膜上皮细胞、慢性胆囊炎上皮细胞与胆囊癌SGC996细胞YAP1 mRNA及蛋白表达差异情况。使用siRNA干扰胆囊癌SGC996细胞中YAP1的表达，并采用RT-PCR法和Western blot法检测EMT相关蛋白表达，MTT法检测细胞活力，流式细胞术检测细胞周期。结果免疫组化显示YAP1在胆囊癌、胆囊炎／胆囊结石组和正常对照组胆囊黏膜中的表达率分别为87．0％（87／100）、56．4％（57／101）和5．0％（1／20）（P〈0．01）。蛋白印迹显示YAP1蛋白在胆囊癌组织及细胞中表达明显高于正常组织及细胞，RT-PCR显示胆囊癌细胞YAP1mRNA水平为正常胆囊黏膜上皮细胞的（12．5±1．2）倍（P〈0．05）。siRNA干扰SGC996细胞中YAPl表达后EMT相关蛋白E．cad、B—catenin、MMP2、FN、Vimentin等表达下降，细胞活力下降，细胞周期被阻滞在S期。结论YAPl与胆囊癌细胞增殖及迁移密切相关，可能通过EMT途径促进肿瘤进展。Objective To investigate the expressions of Yes-associated protein-1 （YAP1） in gall- bladder mucosal epithelium of normal persons, in patients with simple/calculous cholecystitis, and in patients with gallbladder carcinoma; and to study the mechanism of YAP1 in gallbladder carcinoma develop- ment. Methods Immunohistochemistry was used to detect the expression and distribution of YAP1 protein in 50 persons with normal gallbladder, 101 patients with simple cholecystitis/calculous cholecystitis and 100 patients with gallbladder carcinoma. RT-PCR and western-blot were used to detect the mRNA and protein levels of YAP1 in normal and malignant gallbladder mucosal epithelium cells, siRNA was used to shut down the expression of YAP1 in SGC996 cells. MTT was used to test cell vitality. Flow cytometry was used to measure cell cycle. Results Immunohistochemistry revealed the expression rates of YAP1 in the gallbladder carcinoma group, the cholecystitis/gallstone group and the control group to be 87.0% （87/100） , 56.4% （57/101） and 5.0% （1/20） , respectively （P 〈0.01 ）. The YAP1 protein levels were higher in gallbladder carcinoma tissues and cells when compared to normal tissues and cells. RT-PCR showed the mRNA levels of gallbladder carcinoma cells to be 12.5 ± 1.2 times of normal gallbladder mucosal epithelial cells （P 〈 0. 05）. After using siRNA to shut down the YAP1 expression, EMT associated proteins were down-regulated, cell vitality was decreased, and cell cycle was arrested in the S-phase. Conclusions YAP1 is closely related to cell proliferation and metastasis of gallbladder carcinoma. It may promote tumor progression through epithelial-mesenchymal transition.

关 键 词：原发性胆囊癌 Yes相关蛋白1 上皮间质转化 肿瘤进展 

分 类 号：R735.8[医药卫生—肿瘤]