CAJ | 학술논문

目的探讨在人胆囊癌中骨桥蛋白(OPN)的临床意义及其促进转移复发的机制.方法利用实时定量聚合酶链反应(Real-time PCR)技术分析OPN在15例胆囊癌及其对照组织中基因水平的表达;在人胆囊癌细胞株中通过细胞增殖、侵袭等功能实验分析OPN诱导胆囊癌细胞上皮-间充质转化(EMT)促进转移复发的具体机制.结果 OPN在人胆囊癌中较胆囊炎组织表达上调(P＜0.01).克隆形成实验结果显示pWPI-OPN转染组细胞克隆形成数为(59.9±10.3)个,显著高于对照组的(23.7±9.2)个,差异有统计学意义(P＜0.05).在体外实验显示慢病毒转染的OPN高表达胆囊癌细胞株GBC-SD发生EMT,其表现为间质化表型神经钙黏素(N-cadherin)和波形蛋白(Vimentin)上调和上皮化表型上皮钙黏素(E-cadherin)、紧密连接蛋白(ZO-1)的表达下调.同时,它能促进胆囊癌细胞运动和侵袭,过表达OPN组及对照组发生迁移的细胞数分别为[(369.3±23.4)、(84.7±18.7)个,P＜0.01].结论 OPN在胆囊癌组织中高表达与其进展有着直接的联系,其具体机制可能是通过诱导EMT促进胆囊癌的转移复发.
목적 탐토재인담낭암중골교단백(OPN)적림상의의급기촉진전이복발적궤제.방법 이용실시정량취합매련반응(Real-time PCR)기술분석OPN재15례담낭암급기대조조직중기인수평적표체;재인담낭암세포주중통과세포증식、침습등공능실험분석OPN유도담낭암세포상피-간충질전화(EMT)촉진전이복발적구체궤제.결과 OPN재인담낭암중교담낭염조직표체상조(P＜0.01).극륭형성실험결과현시pWPI-OPN전염조세포극륭형성수위(59.9±10.3)개,현저고우대조조적(23.7±9.2)개,차이유통계학의의(P＜0.05).재체외실험현시만병독전염적OPN고표체담낭암세포주GBC-SD발생EMT,기표현위간질화표형신경개점소(N-cadherin)화파형단백(Vimentin)상조화상피화표형상피개점소(E-cadherin)、긴밀련접단백(ZO-1)적표체하조.동시,타능촉진담낭암세포운동화침습,과표체OPN조급대조조발생천이적세포수분별위[(369.3±23.4)、(84.7±18.7)개,P＜0.01].결론 OPN재담낭암조직중고표체여기진전유착직접적련계,기구체궤제가능시통과유도EMT촉진담낭암적전이복발.
Objective To investigate the association between osteopontin (OPN) and the epithelial-mesenchymal transition (EMT) in gallbladder cancer (GBC) and to elucidate the underlying signaling pathway.Methods We evaluated the clinical significance of OPN in 15 GBC patients using real-time quantitative polymerase chain reaction (Real-time PCR).We also investigated the mechanisms of OPN mediating GBC EMT to promote metastasis in GBC cell lines in vitro.Results OPN was up-regulated in the human GBC tissues.The expression of OPN in human GBC tissues was significantly higher than in the cholecystitis tissues (P ＜ 0.01).Clone formation experiment results showed cell clone formation number (59.9 ± 10.3) in pWPI-OPN transfection group was significantly greater than that in control group (23.7 ± 9.2) (P ＜ 0.05).In vitro,the characteristics of EMT were induced by lentivirus transduction of OPN into GBC-SD cells,including the down-regulation of E-cadherin and ZO-1,and the concomitant up-regulation of N-cadherin and vimentin,whereas the ectopic expression of OPN in the GBC cell line promoted cell motility and invasiveness.The number of migrating cells in OPN over-expression group and control group was 369.3 ±23.4 and 84.7 ± 18.7 respectively (P ＜0.01).The further functional studies revealed that OPN regulated the activation of ZEB1,which subsequently regulated EMT signaling pathway.Conclusion Our findings have uncovered a novel role for OPN in GBC metastasis,and provide potential therapeutic target for GBC therapy.