Yes相关蛋白和转录共激活因子PDZ结合基序表达在甲状腺乳头状癌发生发展过程中的作用  

Role of Yes-related proteins and transcriptional co-activator PDZ binding motifs in the development of papillary thyroid carcinoma

作　　者：叶明 张丽婷 吴文艺[1] 林建清[1] 史海鸿[1] 吴鑫泉 余艺煌 丁明骥 邱伟刚 Ye Ming;Zhang Liting;Wu Wenyi;Lin Jianqing;Shi Haihong;Wu Xinquan;Yu Yihuang;Ding Mingji;Qiu Weigang(Department of Thyroid and Breast Surgery,the Second Affiliated Hospital of Fujian Medical University,Quanzhou 362000,China;Department of Endocrinology,the 910th Hospital of the People’s Liberation Army,Quanzhou 362000,China)

机构地区：[1]福建医科大学附属第二临床医学院甲状腺乳腺外科,泉州362000 [2]联勤保障部队第九一〇医院内分泌科,泉州362000

出　　处：《中华实验外科杂志》2022年第4期644-648,共5页Chinese Journal of Experimental Surgery

基　　金：福建省自然科学基金(2021J01251);泉州市科技局高层次人才创业创新项目(2017Z017)。

摘　　要：目的探究Yes相关蛋白(YAP)和转录共激活因子PDZ结合基序(TAZ)在甲状腺乳头癌发生发展过程中的作用。方法用特异性小干扰RNA(siRNA)转染人甲状腺乳头状癌细胞系(TPC-1)使其YAP或TAZ表达水平下调,转染NC为阴性对照组,YAP-si、TAZ-si和TAZ-si+YAP-si为实验组。用实时荧光定量聚合酶链反应(RT-qPCR)、蛋白质印迹法(Western blot)检测转染后TPC-1中YAP和TAZ的表达水平,分别采用细胞计数试剂盒(CCK-8)、流式细胞仪、β-半乳糖苷酶染色法、Transwell实验检测YAP或(和)TAZ低表达对TPC-1增殖、凋亡、衰老、迁移、侵袭的影响。将小鼠随机分成5组,每组6只,将转染后的上述TPC-1接种至裸鼠背部皮下组织(接种体积为0.1 ml/只,接种数量为1×106/裸鼠)。观察并分析在体内YAP或(和)TAZ低表达对肿瘤生长的影响。两样本比较采用t检验。结果YAP-si组TPC-1增殖、迁移、侵袭低于NC组(0.327±0.015比0.361±0.011、0.397±0.021比0.450±0.017、0.560±0.030比0.760±0.036,t=3.188、3.411、7.385,P<0.05;195.000±2.000比361.667±6.429,t=42.875,P<0.01;108.000±8.660比205.333±7.234,t=14.940,P<0.01),衰老阳性、总凋亡率高于NC组[0.185±0.002比0.094±0.001,t=-53.276,P<0.01;(26.367±3.250)%比(7.473±0.729)%,t=-9.824,P<0.01];TAZ-si组TPC-1增殖、迁移、侵袭低于NC组(0.317±0.015比0.361±0.011、0.400±0.010比0.450±0.017、0.537±0.021比0.760±0.036,t=4.056、4.330、9.291,P<0.05;199.677±8.326比361.667±6.429,t=26.673,P<0.01;132.000±13.454比205.333±7.234,t=8.315,P<0.01),衰老阳性、总凋亡率高于NC组[0.183±0.002比0.094±0.001,t=-59.719,P<0.01;(23.367±3.156)%比(7.473±0.729)%,t=-8.479,P<0.01]。YAP-si组肿瘤的体积、重量小于NC组(0.566±0.056比1.021±0.125、2.778±0.257比5.324±0.362、19.877±1.660比31.171±1.962、48.583±3.668比86.441±4.158、80.619±1.243比194.014±9.460、137.865±3.430比331.738±10.514,t=8.127、14.052、10.758、16.726、29.112、42.952,P<0.01;0.730±0.009比1.232±0.015,t=69.Objective To investigate the role of Yes-associated protein(YAP)and transcriptional co-activator PDZ binding motif(TAZ)in the development of papillary thyroid carcinoma.Methods Human papillary thyroid cancer cell line(TPC-1)was transfected with specific small interfering RNA(siRNA)to down-regulate the expression levels of YAP or TAZ.The negative control group was transfected with negative control(NC),and the experimental groups with YAP-si,TAZ-si and TAZ-si+YAP-si respectively.The expression levels of YAP and TAZ in TPC-1 after transfection were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and Western blotting,and the effects of low expression of YAP or(and)TAZ on the proliferation,apoptosis,senescence,migration and invasion of TPC-1 were nalyzed.Mice were randomly divided into 5 groups of 6 mice each,and the transfected TPC-1 mentioned above was inoculated into the subcutaneous tissue of the back of nude mice(inoculation volume:0.1 ml/each;inoculation quantity:1×106/nude mouse).The effect of low expression of YAP or(and)TAZ on tumor growth in vivo was observed and analyzed.Two samples were compared by t-test.Results The proliferation,migration and invasion of TPC-1 cells in the YAP-si group were significantly reduced as compared with those in the NC group(0.327±0.015 vs.0.361±0.011,0.397±0.021 vs.0.450±0.017,0.560±0.030 vs.0.760±0.036,t=3.188,3.411,7.385,P<0.05,and 195.000±2.000 vs.361.667±6.429,t=42.875,P<0.01;108.000±8.660 vs.205.333±7.234,t=14.940,P<0.01).The positive senescence and total apoptosis rate in YAP-si group was higher than in NC group[(0.185±0.002 vs.0.094±0.001,t=-53.276,P<0.01;(26.367±3.250)%vs.(7.473±0.729)%,t=-9.824,P<0.01].The proliferation,migration and invasion of TPC-1 cells in the TAZ-si group were significantly reduced as compared with those in the NC group(0.317±0.015 vs.0.361±0.011,0.400±0.010 vs.0.450±0.017,0.537±0.021 vs.0.760±0.036,t=4.056,4.330,9.291,P<0.05;199.677±8.326 vs.361.667±6.429,t=26.673,P<0.01;132.000±13.454 vs.

关 键 词：甲状腺癌 增殖 迁移 侵袭 

分 类 号：R736.1[医药卫生—肿瘤]