胰腺癌肝转移与循环肿瘤细胞上皮-间质转化表型的关系  

Relationship between liver metastasis and epithelial-mesenchymal transition phenotype of circulating tumor cells in patients with pancreatic cancer

作　　者：祝鹏[1] 刘慧颖[2] 姚骏[1] 柴云笙 徐楷[1] 孙延平[1] 胡志前[1] 王伟军[1] 

机构地区：[1]第二军医大学附属长征医院普通外科,上海200003 [2]第二军医大学附属东方肝胆外科医院分子肿瘤实验室,上海200438

出　　处：《肿瘤》2015年第8期874-882,共9页Tumor

摘　　要：目的 :研究循环肿瘤细胞(circulating tumor cells,CTCs)的上皮-间质转化(epithelial-mesenchymal transition,EMT)表型和胰腺癌肝转移的关系。方法 :收集2010年1月—2013年12月第二军医大学附属长征医院普外科收治、术前影像学检查未发现肝转移且术后病理确诊为胰腺癌的患者,术后随访1年,根据随访期内有无肝转移将患者分为肝转移组和无肝转移组,每组随机选择50例患者。所有患者在术前均采集外周血样本,同时收集10名健康志愿者的外周血作为对照;然后采用免疫磁珠法富集并分离外周血标本中的CTCs,并用氨甲酰磷酸合成酶1(carbamoyl-phosphate synthetase 1,CPS1)抗体的免疫荧光法对CTCs进行鉴定。采用免疫荧光法检测各组患者外周血标本中EMT标志物Snail和Vimentin的表达情况,单因素和多因素分析这些标志物和胰腺癌肝转移的关系。结果 :10名健康志愿者的外周血标本中,CPS1、Snail和Vimentin均无表达;在50例胰腺癌肝转移组患者的外周血标本中,Snail阳性表达41例(82%),Vimentin阳性表达36例(72%);在50例胰腺癌无肝转移组患者的外周血标本中,Snail阳性表达10例(20%),Vimentin阳性表达6例(12%)。肝转移组的Snail和Vimentin阳性表达率均明显高于无肝转移组(χ2=38.455,P<0.01;χ2=36.946,P<0.01)。肝转移组中Snail和Vimentin的表达水平之间存在正相关性(r=0.657,P<0.01)。单因素分析显示,术前糖类抗原19-9(carbohydrate antigen 19-9,CA19-9)水平>37 k U/L、肿瘤直径≥4 cm、胰头肿瘤、血管浸润、Snail表达阳性和Vimentin表达阳性均与胰腺癌肝转移相关(P值均<0.05);多因素分析显示,术前CA19-9>37 k U/L(β=3.411,P<0.01)、Snail表达阳性(β=2.882,P<0.01)和Vimentin表达阳性(β=3.361,P<0.01)是胰腺癌肝转移的独立危险因素。结论 :胰腺癌患者外周血标本中CTCs-EMT表型标志物Snail和Vimentin的表达与其发生肝转移有密切关系。Objective： To study the relationship between epithelial-mesenchymal transition （EMT） phenotype of circulating tumor cells （CTCs） and liver metastasis of pancreatic cancer.Methods： The patients who were found without liver metastasis by preoperative imaging examination but confirmed with pancreatic cancer by postoperative pathological diagnosis were recruited in this study from January 2010 to December 2013 in Department of General Surgery, Changzheng Hospital, Second Military Medical University. All patients were followed up for 1 year after surgical operation, then divided into liver metastasis group and non- liver metastasis group according to whether the patient had liver metastasis or not during the follow-up. In each group, 50 patients were randomly selected. The peripheral blood samples from all patients were collected before surgery, and at the same time, the peripheral blood samples from 10 healthy volunteers were collected as the control. Then the CTCs were enriched and isolated from peripheral blood samples using immunomagnetic method, and identified by immunofluorescence method with carbamoyl-phosphate synthetase 1 （CPS1） antibody. The expressions of EMT markers Snail and Vimentin in blood samples were detected by immunofluorescence method. The relationships between these EMT markers and liver metastasis of pancreatic cancer were analyzed by univariate and multivariate analyses.Results： CPS1, Snail and Vimentin were not expressed in blood samples from 10 healthy volunteers. In peripheral blood samples from 50 pancreatic cancer patients with liver metastasis, the positive expression rates of Snail and Vimentin were 82% （41） and 72% （36）, respectively; while in non-liver metastasis group, the positive expression rates of Snail and Vimentin were 20% （I0） and 12% （6）, respectively. The positive expression rates of Snail and Vimentin in liver metastasis group were higher than those in non-liver metastasis group χ2 = 38.455, P 〈 0.01;χ2 = 36.946, P 〈 0.01）. There was

关 键 词：胰腺肿瘤 肿瘤转移 肝 肿瘤循环细胞 VIMENTIN SNAIL 上皮-间质转化 

分 类 号：R735.9[医药卫生—肿瘤]